Method of treating ocular conditions by administering an antibody that activates Tie2 and binds VEGF

ABSTRACT

The disclosure provides compositions comprising multi-specific compounds, including a compound that targets a phosphatase and a receptor tyrosine kinase agonist. Also provided are methods for the treatment of conditions associated with angiogenesis, comprising administering a multi-specific compound that targets a phosphatase and a receptor tyrosine kinase agonist.

CROSS-REFERENCE

This application is a continuation of U.S. application Ser. No.16/579,078, filed Sep. 23, 2019, now U.S. Pat. No. 10,894,824, whichclaims the benefit of U.S. Provisional Application No. 62/735,331, filedSep. 24, 2018, and U.S. Provisional Application No. 62/832,461, filedApr. 11, 2019, each of which is incorporated herein by reference in itsentirety.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format via EFS-Web and is herebyincorporated by reference in its entirety. Said ASCII copy, created Dec.7, 2020, is named 45725729301_SL.TXT and is 344,503 bytes in size.

BACKGROUND

Individual compounds with the ability to modulate distinct targets canbe combined to generate multispecific compounds. Such multispecificcompounds can have advantages over the parent compounds administeredindividually. These advantages can include, for example, a simplerdosing regimen, longer half-life within a subject, or the ability tobind targets in close proximity.

INCORPORATION BY REFERENCE

Each patent, publication, and non-patent literature cited in theapplication is hereby incorporated by reference in its entirety as ifeach was incorporated by reference individually.

SUMMARY

In some embodiments, the disclosure provides a compound comprising: (a)a first domain, wherein the first domain modulates a phosphatase,wherein the phosphatase modulates Tie2; and (b) a second domain thatspecifically binds a receptor tyrosine kinase agonist.

In some embodiments, the disclosure provides a method of treating acondition in a subject in need thereof, the method comprisingadministering to the subject a therapeutically-effective amount of acompound disclosed herein.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 : Schematic of the basic four chain antibody unit. Light chainsequences are represented by “SEQ A”. Heavy chain sequences arerepresented by “SEQ B”. N and C denote N- and C-termini, respectively.—S—S— denotes disulfide bonds.

FIG. 2 : Schematic of a tetravalent, bispecific antibody with sequencesappended to the heavy chain C-termini. Light chain sequences of the IgGisotype antibody unit are represented by “SEQ A”. Heavy chain sequencesof the IgG isotype antibody unit are represented by “SEQ B”. Linkersequences are represented by “SEQ C”. Appended antigen binding domainsequences are represented by “SEQ D”. N and C denote N- and C-termini,respectively. —S—S— denotes disulfide bonds.

FIG. 3 : Schematic of a tetravalent, bispecific antibody with sequencesappended to the light chain C-termini. Light chain sequences of the IgGisotype antibody unit are represented by “SEQ A”. Heavy chain sequencesof the IgG isotype antibody unit are represented by “SEQ B”. Linkersequences are represented by “SEQ C”. Appended antigen binding domainsequences are represented by “SEQ D”. N and C denote N- and C-termini,respectively. —S—S— denotes disulfide bonds.

FIG. 4 : Schematic of a hexavalent, bispecific antibody with sequencesappended to the heavy chain and light chain C-termini. Light chainsequences of the IgG isotype antibody unit are represented by “SEQ A”.Heavy chain sequences of the IgG isotype antibody unit are representedby “SEQ B”. Linker sequences are represented by “SEQ C” and “SEQ E”.Appended antigen binding domain sequences are represented by “SEQ D”. Nand C denote N- and C-termini, respectively. —S—S— denotes disulfidebonds.

FIG. 5 : Schematic of a hexavalent, trispecific antibody with sequencesappended to the heavy chain and light chain C-termini. Light chainsequences of the IgG isotype antibody unit are represented by “SEQ A”.Heavy chain sequences of the IgG isotype antibody unit are representedby “SEQ B”. Linker sequences are represented by “SEQ C” and “SEQ E”.Appended antigen binding domain sequences are represented by “SEQ D” and“SEQ F”. N and C denote N- and C-termini, respectively. —S—S— denotesdisulfide bonds.

FIG. 6 : Schematic of a bivalent, bispecific antibody with two differentheavy chain sequences and two different light chain sequences. Lightchain sequences are represented by “SEQ A” and “SEQ D”. Heavy chainsequences are represented by “SEQ B” and “SEQ C”. N and C denote N- andC-termini, respectively. —S—S— denotes disulfide bonds.

FIG. 7 : Schematic of a trivalent, trispecific antibody with twodifferent heavy chain sequences, two different light chain sequences,and a sequence appended to the C-terminus of one light chain. Lightchain sequences of the IgG isotype antibody unit are represented by “SEQA” and “SEQ D”. Heavy chain sequences of the IgG isotype antibody unitare represented by “SEQ B” and “SEQ C”. A linker sequence is representedby “SEQ E”. An appended antigen binding domain sequence is representedby “SEQ F”. N and C denote N- and C-termini, respectively. —S—S— denotesdisulfide bonds.

FIG. 8 : Schematic of a trivalent, trispecific antibody with twodifferent heavy chain sequences, two different light chain sequences,and a sequence appended to the C-terminus of one heavy chain. Lightchain sequences of the IgG isotype antibody unit are represented by “SEQA” and “SEQ D”. Heavy chain sequences of the IgG isotype antibody unitare represented by “SEQ B” and “SEQ C”. A linker sequence is representedby “SEQ E”. An appended antigen binding domain sequence is representedby “SEQ F”. N and C denote N- and C-termini, respectively. —S—S— denotesdisulfide bonds.

FIG. 9 : Schematic of a tetravalent antibody with two different heavychain sequences, two different light chain sequences, and sequencesappended to both light chain C-termini. Light chain sequences of the IgGisotype antibody unit are represented by “SEQ A” and “SEQ D”. Heavychain sequences of the IgG isotype antibody unit are represented by “SEQB” and “SEQ C”. Linker sequences are represented by “SEQ E” and “SEQ F”.Appended antigen binding domain sequences are represented by “SEQ G” and“SEQ H”. N and C denote N- and C-termini, respectively. —S—S— denotesdisulfide bonds.

FIG. 10 : Schematic of a tetravalent antibody with two different heavychain sequences, two different light chain sequences, and sequencesappended to both heavy chain C-termini. Light chain sequences of the IgGisotype antibody unit are represented by “SEQ A” and “SEQ D”. Heavychain sequences of the IgG isotype antibody unit are represented by “SEQB” and “SEQ C”. Linker sequences are represented by “SEQ E” and “SEQ F”.Appended antigen binding domain sequences are represented by “SEQ G” and“SEQ H”. N and C denote N- and C-termini, respectively. —S—S— denotesdisulfide bonds.

FIG. 11 : Schematic of a tetravalent antibody with two different heavychain sequences, two different light chain sequences, and sequencesappended to one heavy chain C-terminus and one light chain C-terminus incis. Light chain sequences of the IgG isotype antibody unit arerepresented by “SEQ A” and “SEQ D”. Heavy chain sequences of the IgGisotype antibody unit are represented by “SEQ B” and “SEQ C”. Linkersequences are represented by “SEQ E” and “SEQ F”. Appended antigenbinding domain sequences are represented by “SEQ G” and “SEQ H”. N and Cdenote N- and C-termini, respectively. —S—S— denotes disulfide bonds.

FIG. 12 : Schematic of a tetravalent antibody with two different heavychain sequences, two different light chain sequences, and sequencesappended to one heavy chain C-terminus and one light chain C-terminus intrans. Light chain sequences of the IgG isotype antibody unit arerepresented by “SEQ A” and “SEQ D”. Heavy chain sequences of the IgGisotype antibody unit are represented by “SEQ B” and “SEQ C”. Linkersequences are represented by “SEQ E” and “SEQ F”. Appended antigenbinding domain sequences are represented by “SEQ G” and “SEQ H”. N and Cdenote N- and C-termini, respectively. —S—S— denotes disulfide bonds.

FIG. 13 : Schematic of a pentavalent antibody with two different heavychain sequences, two different light chain sequences, and sequencesappended to both heavy chain C-termini and one light chain C-terminus.Light chain sequences of the IgG isotype antibody unit are representedby “SEQ A” and “SEQ D”. Heavy chain sequences of the IgG isotypeantibody unit are represented by “SEQ B” and “SEQ C”. Linker sequencesare represented by “SEQ E”, “SEQ F”, and “SEQ G”. Appended antigenbinding domain sequences are represented by “SEQ H”, “SEQ I”, and “SEQJ”. N and C denote N- and C-termini, respectively. —S—S— denotesdisulfide bonds.

FIG. 14 : Schematic of a pentavalent antibody with two different heavychain sequences, two different light chain sequences, and sequencesappended to one heavy chain C-terminus and both light chain C-termini.Light chain sequences of the IgG isotype antibody unit are representedby “SEQ A” and “SEQ D”. Heavy chain sequences of the IgG isotypeantibody unit are represented by “SEQ B” and “SEQ C”. Linker sequencesare represented by “SEQ E”, “SEQ F”, and “SEQ G”. Appended antigenbinding domain sequences are represented by “SEQ H”, “SEQ I”, and “SEQJ”. N and C denote N- and C-termini, respectively. —S—S— denotesdisulfide bonds.

FIG. 15 : Schematic of a hexavalent antibody with two different heavychain sequences, two different light chain sequences, and sequencesappended to the heavy chain and light chain C-termini. Light chainsequences of the IgG isotype antibody unit are represented by “SEQ A”and “SEQ D”. Heavy chain sequences of the IgG isotype antibody unit arerepresented by “SEQ B” and “SEQ C”. Linker sequences are represented by“SEQ E”, “SEQ F”, “SEQ G”, and “SEQ H”. Appended antigen binding domainsequences are represented by “SEQ I”, “SEQ J”, “SEQ K” and “SEQ L”. Nand C denote N- and C-termini, respectively. —S—S— denotes disulfidebonds.

FIG. 16 : ELISA data demonstrating binding of a tetravalent bispecificantibody HC2-AFL:LC1 to HPTP-β (top panel) and VEGF (bottom panel).

FIG. 17 : ELISA data demonstrating binding of a tetravalent bispecificantibody HC2-BRO:LC1 to HPTP-β (top panel) and VEGF (bottom panel).

FIG. 18 : ELISA data testing binding of a tetravalent bispecificantibody HC2-RAN:LC1 to HPTP-β (top panel) and VEGF (bottom panel).

FIG. 19 : Activation of Tie2 in HUVECs treated with tetravalentbispecific antibodies or hexavalent bispecific antibodies comprisingbrolucizumab-derived or aflibercept-derived VEGF-binding domains. Asdemonstrated by immunoprecipitation and western blot, all of the testedbispecific antibodies increased basal Tie2 activation (in the absence ofexogenous Ang1 or Ang2), while basal VEGFR2 phosphorylation (in theabsence of exogenous VEGF) was not affected.

FIG. 20 : Tetravalent bispecific and hexavalent bispecific antibodiescomprising brolucizumab-derived or aflibercept-derived VEGF-bindingdomains enhance Tie2 activation and inhibit VEGFR2 activation in HUVECstreated with Ang1 and VEGF, as demonstrated by immunoprecipitation andwestern blot.

FIG. 21A, FIG. 21B, and FIG. 21C: Tetravalent bispecific and hexavalentbispecific antibodies comprising abicipar-derived or aflibercept-derivedVEGF-binding domains enhance Tie2 activation and inhibit VEGFR2activation in HUVECs, including HUVECs treated with Ang1 and VEGF. FIG.21A provides a western blot. FIG. 21B provides quantification of Tie2activation. FIG. 21C provides quantification of VEGFR2 phosphorylation.

FIG. 22A, FIG. 22B, and FIG. 22C: Hexavalent bispecific antibodiescomprising brolucizumab-derived, aflibercept-derived, orabicipar-derived VEGF-binding domains enhance Tie2 activation andinhibit VEGFR2 activation in HUVECs, including HUVECs treated with Ang1and VEGF. FIG. 22A provides a western blot. FIG. 22B providesquantification of Tie2 activation. FIG. 22C provides quantification ofVEGFR2 phosphorylation.

FIG. 23A and FIG. 23B: Tetravalent bispecific and hexavalent bispecificantibodies comprising brolucizumab-derived, aflibercept-derived, orabicipar-derived VEGF-binding domains enhance Tie2 activation andinhibit VEGFR2 activation in HUVECs treated with Ang1 and VEGF, asdemonstrated by electrochemiluminescence signal quantification. FIG. 23Aprovides quantification of Tie2 activation. FIG. 23B providesquantification of VEGFR2 phosphorylation.

FIG. 24A and FIG. 24B: Hexavalent bispecific antibodies comprisingbrolucizumab-derived, aflibercept-derived, or abicipar-derivedVEGF-binding domains enhance Tie2 activation and inhibit VEGFR2activation in HUVECs treated with Ang1 and VEGF, as demonstrated byelectrochemiluminescence signal quantification. FIG. 24A providesquantification of Tie2 activation. FIG. 24B provides quantification ofVEGFR2 phosphorylation.

FIG. 25A and FIG. 25B: Tetravalent bispecific and hexavalent bispecificantibodies comprising brolucizumab-derived VEGF-binding domains enhanceTie2 activation and inhibit VEGFR2 activation in HUVECs treated withAng1 and VEGF, as demonstrated by electrochemiluminescence signalquantification. FIG. 25A provides quantification of Tie2 activation.FIG. 25B provides quantification of VEGFR2 phosphorylation.

FIG. 26A and FIG. 26B: Tetravalent bispecific and hexavalent bispecificantibodies comprising abicipar-derived or aflibercept-derivedVEGF-binding domains enhance Tie2 activation and inhibit VEGFR2activation in HUVECs treated with Ang1 and VEGF, as demonstrated byelectrochemiluminescence signal quantification. FIG. 26A providesquantification of Tie2 activation. FIG. 26B provides quantification ofVEGFR2 phosphorylation.

FIG. 27A and FIG. 27B: Tetravalent bispecific and hexavalent bispecificantibodies comprising abicipar-derived or aflibercept-derivedVEGF-binding domains enhance Tie2 activation and inhibit VEGFR2activation in HUVECs treated with Ang1 and VEGF, as demonstrated byelectrochemiluminescence signal quantification. FIG. 27A providesquantification of Tie2 activation. FIG. 27B provides quantification ofVEGFR2 phosphorylation.

DETAILED DESCRIPTION

The present disclosure provides compositions and methods for modulatingphosphatases and kinases, for example, receptor tyrosine kinases.Compositions and methods are provided for modulating Tie2, for example,to promote Tie2 phosphorylation, signaling, and/or activation. In someembodiments, the disclosure provides compositions and methods fortargeting a phosphatase that modulates Tie2 signaling. In someembodiments, the phosphatase that modulates Tie2 signaling is humanprotein tyrosine phosphatase-beta (HPTP-β).

Compositions and methods are provided for modulating receptor tyrosinekinases, for example, to reduce receptor tyrosine kinasephosphorylation, signaling, and/or activation. In some embodiments, thedisclosure provides compositions and methods for targeting a receptortyrosine kinase agonist, e.g. vascular endothelial growth factor (VEGF).In some embodiments, the disclosure provides compositions and methodsfor targeting a receptor tyrosine kinase, e.g. a VEGF receptor.

In some embodiments, the present disclosure provides compositions andmethods for targeting human protein tyrosine phosphatase-beta (HPTP-β)or vascular endothelial protein tyrosine phosphatase (VE-PTP or VEPTP),and vascular endothelial growth factor (VEGF). In some embodiments, thepresent disclosure provides multispecific compounds, agents, antibodies,fragments, or derivatives thereof that target HPTP-β (VE-PTP) and VEGF.

The agents disclosed herein can be used for the treatment of disordersthat are characterized by, for example, vascular instability,angiogenesis, neovascularization, vascular leakage, and/or edema. Theagents disclosed herein can be used for the treatment of, for example,vascular disorders, ocular disorders, cancers, renal disorders, andcomplications of diabetes.

HPTP-β/VE-PTP, Tie2, and Vascular Stability

HPTP-β is a member of the receptor-like family of the protein tyrosinephosphatases (PTPases). HPTP-β is a transmembrane protein foundprimarily in vascular endothelial cells that displays structural andfunctional similarity to cell adhesion molecules. Orthologues of HPTP-βare found in various species including, for example, zebrafish, chicken,dog, mouse, marmoset, and monkey. The orthologues are generally referredto as vascular endothelial protein tyrosine phosphatase (VE-PTP). HPTP-β(VE-PTP) can influence vascular stability through effects onTie2-mediated signaling.

Tie2 (tyrosine kinase with immunoglobulin and epidermal growth factorhomology domains 2) is a membrane receptor tyrosine kinase expressedprimarily in vascular endothelial cells. Upstream factors can regulateTie2 phosphorylation, influencing downstream signaling and vascularstabilization. Non-limiting examples of such factors includeangiopoietin 1 (Ang1/Angpt1), angiopoietin 2 (Ang2/Angpt2), and HPTP-β(VE-PTP).

Ang1 is an agonist of Tie2. Binding of Ang1 to Tie2 promotes receptorphosphorylation and downstream signaling to induce vascularstabilization through highly organized angiogenesis, tightening ofendothelial cell junctions, enhancement of endothelial viability,reduction of endothelial inflammation, and improvement of endothelialfunction.

Ang2 acts in a context-dependent antagonist or agonist of Tie2. Duringangiogenesis, Ang2 acts as a negative regulator of Ang1-Tie2 signaling.

HPTP-β (VE-PTP) is a phosphatase that can modulate Tie2 signaling.HPTP-β (VE-PTP) can dephosphorylate the Tie2 receptor. Underphysiological conditions, HPTP-β (VE-PTP) regulates the duration of Tie2phosphorylation. Inhibition of HPTP-β (VE-PTP), therefore, can result inincreased Tie2 phosphorylation, increased Tie2-mediated signaling, andenhanced vascular stability. Inhibitors of HPTP-β (VE-PTP) are Tie2activators. For example, a compound, inhibitor, antibody, antibodyfragment, variant, or derivative thereof that binds HPTP-β (VE-PTP) canpromote Tie2 phosphorylation, thereby activating Tie2 downstreamsignaling, and promoting vascular stability.

By the process described above, HPTP-β (VE-PTP) activity can contributeto, for example, disorders that are characterized by vascularinstability, angiogenesis, neovascularization, vascular leakage, and/oredema. For example, HPTP-β (VE-PTP) activity can contribute to vasculardisorders, ocular disorders, cancers, renal disorders, complications ofdiabetes, and other disorders. Inhibition of HPTP-β (VE-PTP) activitycan reduce such disorders.

VEGF and Vascular Stability

Vascular endothelial growth factors (VEGFs) are primarily found inendothelial cells, and are implicated in pathological neovascularizationin a number of diseases. The VEGFs are members of the cystine-knotgrowth factor superfamily, the PDGF family, and the VEGF family. TheVEGFs can act as pro-angiogenic factors. The VEGF family consists ofVEGF-A, VEGF-B, VEGF-C, VEGF-D and placental growth factor (PGF). NineVEGF-A isoforms exist: VEGF₁₂₁, VEGF₁₄₅, VEGF₁₄₈, VEGF₁₆₂, VEGF₁₆₅,VEGF_(165b), VEGF₁₃, VEGF₁₈₉, and VEGF₂₀₆.

VEGF is a hypoxia-regulated gene, and VEGF levels are increased inhypoxic or ischemic conditions. VEGF is an agonist of VEGF receptors(VEGFRs). VEGFRs are receptor tyrosine kinases; binding of VEGF to aVEGFR can result in phosphorylation of the receptor, and subsequently ofdownstream signal transducers. VEGFR-mediated signaling can result inaberrant vasculogenesis, angiogenesis, and permeabilization of bloodvessels, contributing to pathologic vascular instability. Thus,inhibition of VEGF can result in decreased VEGFR-mediated signaling andenhanced vascular stability. For example, an inhibitor, antibody,antibody fragment, variant, or derivative thereof that binds VEGF canreduce VEGFR ligation, thereby reducing VEGFR-mediated signaling, andpromoting vascular stability. Non-limiting examples of agents that bindVEGF include aflibercept (Eylea®), a recombinant protein comprising theVEGF-binding portions of human VEGF receptors 1 and 2 fused to the Fcportion of human IgG1; brolucizumab, a humanized single-chain antibodyfragment (scFv); RTH258, a humanized single-chain antibody fragment(scFv); ranibizumab (Lucentis®), a humanized monoclonal antibodyfragment (Fab); bevacizumab (Avastin®), a humanized monoclonal antibody;conbercept, a recombinant fusion protein comprising extracellulardomains from VEGF receptors 1 and 2 fused to the Fc portion of humanIgG1; Abicipar, a designed ankyrin repeat protein (DARPin); MP0112, aDARPin; MP0250, a DARPin; CT-322, an adnectin; and PRS-050, ananticalin.

By the process described above, VEGF can contribute to, for example,disorders that are characterized by vascular instability, angiogenesis,neovascularization, vascular leakage, and/or edema. For example, VEGFcan contribute to vascular disorders, ocular disorders, cancers, renaldisorders, complications of diabetes, and other disorders. For example,ischemia in the eye can lead to increased VEGF production, resulting invascular leakage and pathological neovascularization in the retina.Inhibition of VEGFR-mediated signaling can reduce such disorders.

Receptor Tyrosine Kinases (RTKs) and Receptor Tyrosine Kinase Agonists

Receptor tyrosine kinases (RTKs) are cell surface receptors thatparticipate in the regulation of cell growth, differentiation, andsurvival. Binding of an agonist to a RTK can cause neighboring RTKs toassociate with each other, forming dimers. Dimerization can causecross-phosphorylation—each RTK in the dimer phosphorylates multipletyrosine residues on the other RTK. Once cross-phosphorylated, thecytoplasmic tails of the RTKs can initiate signal transduction pathways,for example, by serving as docking platforms for various intracellularproteins. RTK signaling can lead to changes of gene transcription andexpression in a cell.

Non-limiting examples of RTKs include AATK, AATYK, AATYK1, AATYK2, ACH,ALK, ARK, AXL, BDB, BDB1, BEK, BFGFR, BREK, Brt, CAK, CCK4, CD115,CD117, CD135, CD136, CD140a, CD140b, CD167, CD202b, CD220, CD221, CD246,CD309, CD331, CD332, CD333, CD334, CDHF12, CDHR16, CDw136, CEK, CEK2,CEK3, c-Eyk, CFD1, C-FMS, C-Kit, cprk, c-ros-1, CSF1R, CSFR, D3S3195,DDR1, DDR2, DFNB97, DKFZp761P1010, Dtk, ECT1, EDDR1, EGFR, EphA10,EphA1-8. EphB1, EphB2, EphB3, EphB4, EphB6, ErbB2, ErbB3, ErbB4, Etk-2,FGFR1, FGFR2. FGFR3. FGFR4, FLG, FLK1, FLK2, FLT, FLT1, FLT2, FLT3,FLT4, FMS, GAS9, H2, H3, H4, H5, HGFR, HSCR1, IGF1R, IGFIR, IGFR, INSR,INSRR, IRR, JKT5A, JTK11, JTK12, JTK13, JTK14, JTK2, JTK4, JTK5, JWS,KAL2, KDR, KGFR, KIAA0641, KIAA1079, KIAA1883, KIT, KPI2, K-SAM, LMR1,LMR2, LMR3, LMTK1, LMTK2, LMTK3, LTK, MCF3, MEN2A, MEN2B, Mer, MERTK,MET, MGC18216, MST1R, MTC, MTC1, MTRK1, MuSK, NEP, NOK, N-SAM, NTRK2,NTRK3, NTRK4, NTRKR1, NTRKR2, PBT, PCL, PDGFR, PDGFR1, PDGFR2, PDGFRA,PDGFR-alpha, PDGFRB, PDGFR-beta, PPP1R100, PPP1R101, PPP1R77, PTC,PTK3A, PTK7, PTK8, RCCP2, Rek, RET, RET51, RON, ROR1, ROR2, ROS, ROS1,RP38, RSE, RTK6, RYK, Ryk, RYK1, SCFR, Sky, STK, STYK1, SuRTK106, TEK,TIE1, TIE2, Tif, TK14, TK25, TKT, TRK, TrkA, TrkB, TrkC, TYK1, TYKLM3,TYRO10, Tyro12, Tyro3, Tyro7, UFO, VEGFR, VEGFR1, VEGFR2, VEGFR3, VMCM,and VMCM1.

Non-limiting examples of RTK agonists include VEGF, Ang1, Ang2, BDNF,EGF, FGF, HGF, IGF, insulin, MSP, NGF, NT-3, and PDGF.

Antibodies and Antigen-Binding Compounds.

The basic four chain antibody unit comprises two identical heavy chain(H) polypeptide sequences and two identical light chain (L) polypeptidesequences. Each of the heavy chains can comprise one N-terminal variable(V_(H)) region and three or four C-terminal constant (C_(H)1, C_(H)2,C_(H)3, and C_(H)4) regions. Each of the light chains can comprise oneN-terminal variable (V_(L)) region and one C-terminal constant (C_(L))region. The light chain variable region is aligned with the heavy chainvariable region and the light chain constant region is aligned withheavy chain constant region C_(H1). The pairing of a heavy chainvariable region and light chain variable region together forms a singleantigen-binding site. Each light chain is linked to a heavy chain by onecovalent disulfide bond. The two heavy chains are linked to each otherby one or more disulfide bonds depending on the heavy chain isotype.Each heavy and light chain also comprises regularly-spaced intrachaindisulfide bridges. The C-terminal constant regions of the heavy chainscomprise the Fc region of the antibody, which mediate effectorfunctions, for example, through interactions with Fc receptors orcomplement proteins. FIG. 1 provides a simple representative schematicbasic four chain antibody unit; light chain sequences are represented by“SEQ A”. Heavy chain sequences are represented by “SEQ B”, —S—S— denotesdisulfide bonds, N and C denote N- and C-termini, respectively.

The light chain can be designated kappa or lambda based on the aminoacid sequence of the constant region. The heavy chain can be designatedalpha, delta, epsilon, gamma, or mu based on the amino acid sequence ofthe constant region. Antibodies are categorized into five immunoglobulinclasses, or isotypes, based on the heavy chain. IgA comprises alphaheavy chains, IgD comprises delta heavy chains, IgE comprises epsilonheavy chains, IgG comprises gamma heavy chains, and IgM comprises muheavy chains. Antibodies of the IgG, IgD, and IgE classes comprisemonomers of the four chain unit described above (two heavy and two lightchains), while the IgM and IgA classes can comprise multimers of thefour chain unit. The alpha and gamma classes are further divided intosubclasses on the basis of differences in the sequence and function ofthe heavy chain constant region. Subclasses of IgA and IgG expressed byhumans include IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2.

The constant regions are minimally involved in antigen binding. Rather,the constant regions can mediate various effector functions. DifferentIgG isotypes or subclasses can be associated with different effectorfunctions or therapeutic characteristics, for example, because ofinteractions with different Fc receptors and/or complement proteins.Antibodies comprising Fc regions that engage activating Fc receptorscan, for example, participate in antibody-dependent cell-mediatedcytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP),complement-dependent cytotoxicity (CDC), induction of signaling throughimmunoreceptor tyrosine-based activation motifs (ITAMs), and inductionof cytokine secretion. Antibodies comprising Fc regions that engageinhibitory Fc receptors can, for example, induce signaling throughimmunoreceptor tyrosine-based inhibitory motifs (ITIMs).

Different antibody subclasses comprise different abilities to elicitimmune effector functions. For example, IgG1 and IgG3 can effectivelyrecruit complement to activate CDC, IgG2 elicits minimal ADCC. IgG4 hasa lesser ability to trigger immune effector functions. Modifications tothe constant regions can also affect antibody characteristics, forexample, enhancement or reduction of Fc receptor ligation, enhancementor reduction of ADCC, enhancement or reduction of ADCP, enhancement orreduction of CDC, enhancement or reduction of signaling through ITAMs,enhancement or reduction of cytokine induction, enhancement or reductionof signaling through ITIMs, enhancement or reduction of half-life, orenhancement or reduction of coengagement of antigen with Fc receptors.Modifications can include, for example, amino acid mutations, alteringpost-translational modifications (e.g., glycosylation), combiningdomains from different isotypes or subclasses, or a combination thereof.

A compound or antibody of the disclosure can comprise constant regionsor Fc regions that are selected or modified to provide suitable antibodycharacteristics, for example, suitable characteristics for treating adisease or condition as disclosed herein. In some embodiments, IgG1 canbe used, for example, to promote immune activation effector functions(e.g., ADCC, ADCP, CDC, ITAM signaling, cytokine induction, or acombination thereof for the treatment of a cancer). In some embodiments,IgG4 can be used, for example, in cases where antagonistic properties ofthe antibody in the absence of immune effector functions are desirable(e.g., for treatment of ocular disorders).

Non-limiting examples of antibody modifications and their effects areprovided in TABLE 1.

TABLE 1 Effect Isotype Mutation(s)/modification(s) Enhanced ADCC IgG1F243L/R292P/Y300L/V305I/ P396L Enhanced ADCC IgG1 S239D/I332E EnhancedADCC IgG1 S239D/I332E/A330L Enhanced ADCC IgG1 S298A/E333A/K334AEnhanced ADCC IgG1 In one heavy chain: L234Y/ L235Q/G236W/S239M/H268D/D270E/S298A In the opposing heavy chain: D270E/K326D/A330M/K334EEnhanced ADCP IgG1 G236A/S239D/I332E Enhanced CDC IgG1 K326W/E333SEnhanced CDC IgG1 S267E/H268F/S324T Enhanced CDC IgG1, Combination ofdomains from IgG3 IgG1/IgG3 Enhanced CDC IgG1 E345R/E430G/S440Y Loss ofglycosylation, IgG1 N297A or N297Q or N297G reduced effector functionsReduced effector functions IgG1, L235E IgG4 Reduced effector functionsIgG1 L234A/L235A Reduced effector functions IgG4 F234A/L235A Reducedeffector functions IgG4 F234A/L235A/G237A/P238S Reduced effectorfunctions IgG4 F234A/L235A/ΔG236/ G237A/P238S Reduced effector functionsIgG2, Combination of domains from IgG4 IgG2/IgG4 Reduced effectorfunctions IgG2 H268Q/V309L/A330S/P331S Reduced effector functions IgG2V234A/G237A/P238S/H268A/ V309L/A330S/P331S Reduced effector functionsIgG1 L234A/L235A/G237A/P238S/ H268A/A330S/P331S Increased half-life IgG1M252Y/S254T/T256E Increased half-life IgG1 M428L/N434S Increasedantigen/Fc IgG1 S267E/L328F receptor coengagement Altered antigen/FcIgG1 N325S/L328F receptor coengagement Reduced Fab arm exchange IgG4S228P

The variable (V) regions mediate antigen binding and define thespecificity of a particular antibody for an antigen. The variable regioncomprises relatively invariant sequences called framework regions, andhypervariable regions, which differ considerably in sequence amongantibodies of different binding specificities. The variable region ofeach antibody heavy or light chain comprises four framework regionsseparated by three hypervariable regions. The variable regions of heavyand light chains fold in a manner that brings the hypervariable regionstogether in close proximity to create an antigen binding site. The fourframework regions largely adopt an f3-sheet configuration, while thethree hypervariable regions form loops connecting, and in some casesforming part of, the f3-sheet structure.

Within hypervariable regions are amino acid residues that primarilydetermine the binding specificity of the antibody. Sequences comprisingthese residues are known as complementarity determining regions (CDRs).One antigen binding site of an antibody comprises six CDRs, three in thehypervariable regions of the light chain, and three in the hypervariableregions of the heavy chain. The CDRs in the light chain are designatedL1, L2, and L3, while the CDRs in the heavy chain are designated H1, H2,and H3. CDRs can also be designated LCDR1, LCDR2, LCDR3, HCDR1, HCDR2,and HCDR3, respectively. The contribution of each CDR to antigen bindingvaries among antibodies. CDRs can vary in length. For example, CDRs areoften 5 to 14 residues in length, but CDRs as short as 0 residues or aslong as 25 residues or longer exist.

Several methods are used to predict or designate CDR sequences. Thesemethods can use different numbering systems, for example, becausesequence insertions and deletions are numbered differently.

The Kabat method was developed by aligning a limited number of antibodysequences and determining the positions of the most variable residues.Based on the alignment, a numbering scheme was introduced for residuesin the variable regions. This numbering scheme can be used to determinethe positions marking the beginning and the end of each CDR. Oneiteration of the Kabat numbering system identifies CDRs in the lightchain variable region using the following residue positions: LCDR1around residues 24-34; LCDR2 around residues 50-56; and LCDR3 aroundresidues 89-97. One iteration of the Kabat numbering system identifiesCDRs in the heavy chain variable region using the following residuepositions: HCDR1 around residues 31-35; HCDR2 around residues 50-65; andHCDR3 around residues 95-102.

The Chothia method was developed based on analysis of three dimensionalantibody structures. The analysis determined that hypervariable loopsadopt a restricted set of conformations based on the presence of certainresidues at key positions in CDRs and flanking framework regions. Thismethod uses a similar numbering scheme as the Kabat method, but numbersinsertions and deletions differently. One iteration of the Chothianumbering system identifies CDRs in the light chain variable regionusing the following residue positions: LCDR1 around residues 24-34;LCDR2 around residues 50-56; and LCDR3 around residues 89-97. Oneiteration of the Chothia numbering system identifies CDRs in the heavychain variable region using the following residue positions: HCDR1around residues 26-34; HCDR2 around residues 52-56; and HCDR3 aroundresidues 95-102.

The IMGT method (International ImMunoGeneTics database) was developed byintegrating existing definitions of framework regions and CDRs,structural data, and data from alignment of antibody variable regionsequences. This integration led to the identification of conservedresidues in the framework regions that can be used as reference pointsfor identifying CDRs. Examples of conserved residues in variable regionsinclude cysteine at approximately residue 23 (in framework region 1),tryptophan at approximately residue 41 (in framework region 2), ahydrophobic amino acid at approximately residue 89 (in framework region3), cysteine at approximately residue 104 (in framework region 3), andphenylalanine or tryptophan at approximately residue 118 (in frameworkregion 4). CDRs can be identified in a sequence encoding an antibodyvariable region of interest by using a computational alignment-basedalgorithm.

The IMGT method of numbering consistently assigns the same numbers tothe conserved amino acids, but the lengths of CDRs and framework regionsare permitted to vary. Therefore, IMGT numbering of residues is notnecessarily sequential. The length of CDRs identified by the IMGT methodcan vary. For example, LCDR1 or HCDR1 can be about 5 to about 12 aminoacids, LCDR2 or HCDR2 can be about 0 to about 10 amino acids, and LCDR3or HCDR3 can be about 5 to about 91 amino acids.

The Paratome method was developed based on multiple structuralalignments of available antibody-antigen complexes. The structuralpositions that bind antigen were found to be similar among the examinedantibodies, and antibody sequences from the data set were annotated withAntigen Binding Regions (ABRs, similar to CDRs). ABRs in a querysequence can be identified using a computational tool, which firstaligns the query sequence against antibodies with solvedantibody-antigen structures, then infers the positions of ABRs based onthe alignment. Antibodies with solved structures can also have ABRsidentified using a structural, rather than sequence-based, alignmentmethod.

A subset of residues within CDRs contacts an antigen. These residuesthat contact antigen can be referred to as specificity-determiningresidues (SDRs). However, residues other than SDRs can contribute tobinding activity by helping to maintain the conformation of the bindingsite. The number of SDRs in an antibody can vary based on the size andtype of antigen that is recognized, for example, between 0-14 SDRs canbe found within a CDR. SDRs can be enriched in some residues, such astyrosine, serine, tryptophan, and asparagine.

A monoclonal antibody can be obtained from a population ofsubstantially-homogeneous antibodies, i.e., the individual antibodiescomprising the population are identical except for possiblenaturally-occurring mutations that can be present in minor amounts. Incontrast to polyclonal antibody preparations, which include differentantibodies directed against different epitopes, each monoclonal antibodyis directed against a single epitope.

A compound herein can be a monoclonal antibody, for example, a chimericantibody wherein a portion of the heavy and/or light chain is identicalto or homologous to a corresponding sequence in an antibody derived froma particular species or belonging to a particular antibody class orsubclass, while the remainder of the chain(s) is identical or homologousto a corresponding sequence in an antibody derived from another speciesor belonging to another antibody class or subclass, or anantigen-binding fragment of such an antibody.

An antibody fragment or antigen-binding fragment can comprise a portionof an antibody, for example, the antigen-binding or variable region ofthe intact antibody. Non-limiting examples of antibody fragments includeFab, Fab′, F(ab′)₂, dimers and trimers of Fab conjugates, Fv, scFv,minibodies, dia-, tria-, and tetrabodies, and linear antibodies. Fab andFab′ are antigen-binding fragments that can comprise the V_(H) andC_(H)1 domains of the heavy chain linked to the V_(L) and C_(L) domainsof the light chain via a disulfide bond. A F(ab′)₂ can comprise two Fabor Fab′ that are joined by disulfide bonds. A Fv can comprise the V_(H)and V_(L) domains held together by non-covalent interactions. A scFv(single-chain variable fragment) is a fusion protein that can comprisethe V_(H) and V_(L) domains connected by a peptide linker. Manipulationof the orientation of the V_(H) and V_(L) domains and the linker lengthcan be used to create different forms of molecules that can bemonomeric, dimeric (diabody), trimeric (triabody), or tetrameric(tetrabody). Minibodies are scFv-C_(H)3fusion proteins that assembleinto bivalent dimers.

Non-limiting examples of epitopes include amino acids, sugars, lipids,phosphoryl, and sulfonyl groups. An epitope can have specificthree-dimensional structural characteristics, and/or specific chargecharacteristics. Epitopes can be conformational or linear.

For human administration, monoclonal antibodies generated from non-humanspecies can be further optimized by a humanization process to reduce thelikelihood of immunogenicity while preserving target specificity.Humanization processes involve the incorporation of human DNA to thegenetic sequence of the genes that produce the isolated antibodies. Therecombinant DNA is cloned and expressed in cells for large-scaleproduction of the newly humanized antibodies.

An example of a humanized antibody is a modified chimeric antibody. Achimeric antibody can be generated as described above. The chimericantibody is further mutated outside of the CDRs to substitute non-humansequences in the variable regions with the homologous human sequences.Another example of a humanized antibody is a CDR-grafted antibody, inwhich non-human CDR sequences are introduced into the human heavy andlight chain variable sequences of a human antibody scaffold to replacethe corresponding human CDR sequences.

A humanized antibody can be produced in mammalian cells, bioreactors, ortransgenic animals, such as mouse, chicken, sheep, goat, pig, ormarmoset. The transgenic animal can have a substantial portion of thehuman antibody-producing genome inserted into the genome of the animal.

In addition to antibodies and antibody fragments, other antigen-bindingcompounds can also bind target molecules. Non-limiting examples ofnon-antibody-derived antigen-binding compounds include ankyrin proteins,ankyrin repeat proteins, designed ankyrin repeat proteins (DARPins),affibodies, avimers, adnectins, anticalins, Fynomers, Kunitz domains,knottins, β-hairpin mimetics, and receptors and derivatives thereof,e.g. VEGF receptors, or the VEGF-binding portions of human VEGFreceptors 1 and 2.

Designed ankyrin repeat proteins (DARPins) can be protein scaffoldsbased on ankyrin repeat proteins. A DARPin can comprise one or moreankyrin repeats that comprise a shared sequence and/or structural motif.The individual ankyrin repeats can comprise a shared sequence and/orstructural motif despite comprising mutations, substitutions, additionsand/or deletions when compared to one other. A DARPin can comprise, forexample, about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 ankyrin repeats, ormore. A DARPin can comprise an N-terminal capping repeat, one or moreinternal ankyrin repeats, and a C-terminal capping repeat. Each ankyrinrepeat can comprise framework residues and protein-interaction residues.The framework residues can contribute to structure or folding topology,for example, the structure of an ankyrin repeat or interaction with aneighboring ankyrin repeat. Protein-interaction residues can contributeto binding of a target molecule, for example, via direct interactionwith the target molecule, or by stabilizing directly-interactingresidues in a conformation that allows binding.

Compounds, antibodies, fragments or derivatives thereof, or othercompounds that bind target molecules in this disclosure can bind totargets with a K_(D) of, for example, less than about 500 nM, less thanabout 300 nM, less than about 200 nM, less than about 100 nM, less thanabout 90 nM, less than about 80 nM, less than about 70 nM, less thanabout 60 nM, less than about 50 nM, less than about 40 nM, less thanabout 30 nM, less than about 20 nM, less than about 10 nM, less thanabout 5 nM, less than about 4 nM, less than about 3 nM, less than about2 nM, less than about 1 nM, less than about 900 pM, less than about 800pM, less than about 700 pM, less than about 600 pM, less than about 500pM, less than about 400 pM, less than about 300 pM, less than about 200pM, less than about 100 pM, less than about 90 pM, less than about 80pM, less than about 70 pM, less than about 60 pM, less than about 50 pM,less than about 40 pM, less than about 30 pM, less than about 20 pM,less than about 10 pM, less than about 9 pM, less than about 8 pM, lessthan about 7 pM, less than about 6 pM, less than about 5 pM, less thanabout 4 pM, less than about 3 pM, less than about 2 pM, less than about1 pM, less than about 900 fM, less than about 800 fM, less than about700 fM, less than about 600 fM, less than about 500 fM, less than about400 fM, less than about 300 fM, less than about 200 fM, less than about100 fM, less than about 90 fM, less than about 80 fM, less than about 70fM, less than about 60 fM, less than about 50 fM, less than about 40 fM,less than about 30 fM, less than about 20 fM, or less than about 10 fM.

In some embodiments, a compound, antibody, fragment or derivativesthereof, or other compound that a bind target molecule in thisdisclosure can bind to the target with a K_(D) of, for example, about 10fM to about 500 nM, about 30 fM to about 500 nM, about 30 fM to about400 nM, about 30 fM to about 300 nM, about 30 fM to about 200 nM, about30 fM to about 100 nM, about 30 fM to about 90 nM, about 30 fM to about80 nM, about 30 fM to about 70 nM, about 30 fM to about 60 nM, about 30fM to about 50 nM, about 30 fM to about 40 nM, about 30 fM to about 30nM, about 30 fM to about 20 nM, about 30 fM to about 10 nM, about 30 fMto about 9 nM, about 30 fM to about 8 nM, about 30 fM to about 7 nM,about 30 fM to about 6 nM, about 30 fM to about 5 nM, about 30 fM toabout 4 nM, about 30 fM to about 3 nM, about 30 fM to about 2 nM, about30 fM to about 1 nM, about 30 fM to about 900 pM, about 30 fM to about800 pM, about 30 fM to about 700 pM, about 30 fM to about 600 pM, about30 fM to about 500 pM, about 30 fM to about 400 pM, about 30 fM to about300 pM, about 30 fM to about 200 pM, about 30 fM to about 100 pM, about30 fM to about 90 pM, about 30 fM to about 80 pM, about 30 fM to about70 pM, about 30 fM to about 60 pM, about 30 fM to about 50 pM, about 30fM to about 40 pM, about 30 fM to about 30 pM, about 30 fM to about 20pM, about 30 fM to about 10 pM, about 30 fM to about 1 pM, about 30 fMto about 900 fM, about 30 fM to about 800 fM, about 30 fM to about 700fM, about 30 fM to about 600 fM, about 30 fM to about 500 fM, about 30fM to about 400 fM, about 30 fM to about 300 fM, about 30 fM to about200 fM, about 30 fM to about 100 fM, about 30 fM to about 500 nM, about30 fM to about 400 nM, about 30 fM to about 300 nM, about 30 fM to about200 nM, about 30 fM to about 100 nM, about 30 fM to about 90 nM, about30 fM to about 80 nM, about 30 fM to about 70 nM, about 30 fM to about60 nM, about 30 fM to about 50 nM, about 30 fM to about 40 nM, about 30fM to about 30 nM, about 30 fM to about 20 nM, about 30 fM to about 10nM, about 30 fM to about 9 nM, about 30 fM to about 8 nM, about 30 fM toabout 7 nM, about 30 fM to about 6 nM, about 30 fM to about 5 nM, about30 fM to about 4 nM, about 30 fM to about 3 nM, about 30 fM to about 2nM, about 30 fM to about 1 nM, about 30 fM to about 900 pM, about 30 fMto about 800 pM, about 30 fM to about 700 pM, about 30 fM to about 600pM, about 30 fM to about 500 pM, about 30 fM to about 400 pM, about 30fM to about 300 pM, about 30 fM to about 200 pM, about 30 fM to about100 pM, about 30 fM to about 90 pM, about 30 fM to about 80 pM, about 30fM to about 70 pM, about 30 fM to about 60 pM, about 30 fM to about 50pM, about 30 fM to about 40 pM, about 30 fM to about 30 pM, about 30 fMto about 20 pM, about 30 fM to about 10 pM, about 1 pM to about 500 nM,about 1 pM to about 400 nM, about 1 pM to about 300 nM, about 1 pM toabout 200 nM, about 1 pM to about 100 nM, about 1 pM to about 90 nM,about 1 pM to about 80 nM, about 1 pM to about 70 nM, about 1 pM toabout 60 nM, about 1 pM to about 50 nM, about 1 pM to about 40 nM, about1 pM to about 30 nM, about 1 pM to about 20 nM, about 1 pM to about 10nM, about 1 pM to about 9 nM, about 1 pM to about 8 nM, about 1 pM toabout 7 nM, about 1 pM to about 6 nM, about 1 pM to about 5 nM, about 1pM to about 4 nM, about 1 pM to about 3 nM, about 1 pM to about 2 nM,about 1 pM to about 1 nM, about 1 pM to about 900 pM, about 1 pM toabout 800 pM, about 1 pM to about 700 pM, about 1 pM to about 600 pM,about 1 pM to about 500 pM, about 1 pM to about 400 pM, about 1 pM toabout 300 pM, about 1 pM to about 200 pM, about 1 pM to about 100 pM,about 1 pM to about 90 pM, about 1 pM to about 80 pM, about 1 pM toabout 70 pM, about 1 pM to about 60 pM, about 1 pM to about 50 pM, about1 pM to about 40 pM, about 1 pM to about 30 pM, about 1 pM to about 20pM, about 1 pM to about 10 pM, about 100 pM to about 500 nM, about 100pM to about 400 nM, about 100 pM to about 300 nM, about 100 pM to about200 nM, about 100 pM to about 100 nM, about 100 pM to about 90 nM, about100 pM to about 80 nM, about 100 pM to about 70 nM, about 100 pM toabout 60 nM, about 100 pM to about 50 nM, about 100 pM to about 40 nM,about 100 pM to about 30 nM, about 100 pM to about 20 nM, about 100 pMto about 10 nM, about 100 pM to about 9 nM, about 100 pM to about 8 nM,about 100 pM to about 7 nM, about 100 pM to about 6 nM, about 100 pM toabout 5 nM, about 100 pM to about 4 nM, about 100 pM to about 3 nM,about 100 pM to about 2 nM, about 100 pM to about 1 nM, about 100 pM toabout 900 pM, about 100 pM to about 800 pM, about 100 pM to about 700pM, about 100 pM to about 600 pM, about 100 pM to about 500 pM, about100 pM to about 400 pM, about 100 pM to about 300 pM, or about 100 pM toabout 200 pM.

In some embodiments, a compound, antibody, fragment or derivativesthereof, or other compound of the disclosure can bind HPTP-β (VE-PTP)with a K_(D) of about 500 fM to about 500 pM. In some embodiments, acompound, antibody, fragment or derivatives thereof, or other compoundof the disclosure can bind HPTP-β (VE-PTP) with a K_(D) of about 1 pM toabout 500 pM. In some embodiments, a compound, antibody, fragment orderivatives thereof, or other compound of the disclosure can bind HPTP-β(VE-PTP) with a K_(D) of about 60 pM to about 500 pM. In someembodiments, a compound, antibody, fragment or derivatives thereof, orother compound of the disclosure can bind HPTP-β (VE-PTP) with a K_(D)of about 100 pM to about 500 pM. In some embodiments, a compound,antibody, fragment or derivatives thereof, or other compound of thedisclosure can bind HPTP-β (VE-PTP) with a K_(D) of about 1 pM to about300 pM. In some embodiments, a compound, antibody, fragment orderivatives thereof, or other compound of the disclosure can bind HPTP-β(VE-PTP) with a K_(D) of about 1 pM to about 200 pM. In someembodiments, a compound, antibody, fragment or derivatives thereof, orother compound of the disclosure can bind HPTP-β (VE-PTP) with a K_(D)of about 1 pM to about 120 pM. In some embodiments, a compound,antibody, fragment or derivatives thereof, or other compound of thedisclosure can bind HPTP-β (VE-PTP) with a K_(D) of about 1 pM to about70 pM.

In some embodiments, a compound, antibody, fragment or derivativesthereof, or other compound of the disclosure can bind VEGF with a K_(D)of about 30 fM to about 900 pM. In some embodiments, a compound,antibody, fragment or derivatives thereof, or other compound of thedisclosure can bind VEGF with a K_(D) of about 30 fM to about 600 pM. Insome embodiments, a compound, antibody, fragment or derivatives thereof,or other compound of the disclosure can bind VEGF with a K_(D) of about30 fM to about 200 pM. In some embodiments, a compound, antibody,fragment or derivatives thereof, or other compound of the disclosure canbind VEGF with a K_(D) of about 30 fM to about 30 pM. In someembodiments, a compound, antibody, fragment or derivatives thereof, orother compound of the disclosure can bind VEGF with a K_(D) of about 30fM to about 40 pM. In some embodiments, a compound, antibody, fragmentor derivatives thereof, or other compound of the disclosure can bindVEGF with a K_(D) of about 30 fM to about 1 pM. In some embodiments, acompound, antibody, fragment or derivatives thereof, or other compoundof the disclosure can bind VEGF with a K_(D) of about 30 fM to about 200fM. In some embodiments, a compound, antibody, fragment or derivativesthereof, or other compound of the disclosure can bind VEGF with a K_(D)of about 1 pM to about 900 pM. In some embodiments, a compound,antibody, fragment or derivatives thereof, or other compound of thedisclosure can bind VEGF with a K_(D) of about 1 pM to about 600 pM. Insome embodiments, a compound, antibody, fragment or derivatives thereof,or other compound of the disclosure can bind VEGF with a K_(D) of about1 pM to about 200 pM. In some embodiments, a compound, antibody,fragment or derivatives thereof, or other compound of the disclosure canbind VEGF with a K_(D) of about 1 pM to about 30 pM. In someembodiments, a compound, antibody, fragment or derivatives thereof, orother compound of the disclosure can bind VEGF with a K_(D) of about 1pM to about 40 pM.

In some embodiments, a compound, antibody, fragment or derivativesthereof, or other compound of the disclosure can bind VEGF with a K_(D)of about 30 fM to about 2 pM. In some embodiments, a compound, antibody,fragment or derivatives thereof, or other compound of the disclosure canbind VEGF with a K_(D) of about 35 fM to about 200 fM. In someembodiments, a compound, antibody, fragment or derivatives thereof, orother compound of the disclosure can bind VEGF with a K_(D) of about 100fM to about 2 pM.

In some embodiments, a compound, antibody, fragment or derivativesthereof, or other compound of the disclosure can bind VEGF with a K_(D)of about 20 pM to about 1 nM. In some embodiments, a compound, antibody,fragment or derivatives thereof, or other compound of the disclosure canbind VEGF with a K_(D) of about 20 pM to about 800 pM. In someembodiments, a compound, antibody, fragment or derivatives thereof, orother compound of the disclosure can bind VEGF with a K_(D) of about 20pM to about 350 pM.

In some embodiments, a compound, antibody, fragment or derivativesthereof, or other compound of the disclosure can bind VEGF with a K_(D)of about 30 fM to about 700 pM.

In some embodiments, a compound, antibody, fragment or derivativesthereof, or other compound of the disclosure can bind VEGF with a K_(D)of about 40 fM to about 520 pM. In some embodiments, a compound,antibody, fragment or derivatives thereof, or other compound of thedisclosure can bind VEGF with a K_(D) of about 40 fM to about 110 pM.

Polyvalent, Multispecific Compounds

Antigen-binding compounds can be combined to generate polyvalent and/ormultispecific compounds. Such polyvalent and/or multispecific compoundscan have advantages over the parent compounds administered individually.These advantages can include, for example, a simpler dosing regimen,longer half-life within a subject, and the ability to bind targetantigens in close proximity.

Multispecific antibodies can be produced by a number of methods. In onemethod, monospecific antibodies or derivatives thereof can be chemicallycoupled, for example, via chemical coupling of two IgG antibody unitsinto a conjugate.

In another method, cloning techniques can be used to append additionalantigen binding domain(s) to a conventional IgG antibody or derivativethereof. An additional antigen-binding domain can be, for example, asingle variable domain (sVD), a single-chain variable fragment (scFv), asingle-chain Fab, a peptide, an ankyrin protein, an ankyrin repeatprotein, a designed ankyrin repeat protein (DARPin), an affibody, anavimer, an adnectin, an anticalin, a Fynomer, a Kunitz domain, aknottin, a β-hairpin mimetic, a tetrameric polyethylene oxide clusteredpeptide, a peptide derived from one or more receptors (e.g. VEGFreceptors, or the VEGF-binding portions of human VEGF receptors 1 and2), or a derivative thereof.

The additional antigen-binding domain (e.g., sVD, scFv, single-chainFab, peptide, ankyrin protein, ankyrin repeat protein, DARPin, affibody,avimer, adnectin, anticalin, Fynomer, Kunitz domain, knottin, β-hairpinmimetic, tetrameric polyethylene oxide clustered peptide, or peptidederived from one or more receptors) can be appended to the N orC-terminus of the light chain and/or heavy chain of the IgG or Fab, forexample, via a peptide linker. In some embodiments, a scFv can beappended to the C-termini of the heavy chains of an IgG to provide atetravalent bispecific antibody. A scFv can be appended to the C-terminiof the light chains of an IgG to provide a tetravalent bispecificantibody. A scFv can be appended to the C-termini of the light chainsand the C-termini of the heavy chains of an IgG, to provide a hexavalentbispecific antibody. In some embodiments, a DARPin can be appended tothe C-termini of the heavy chains of an IgG to provide a tetravalentbispecific antibody. A DARPin can be appended to the C-termini of thelight chains of an IgG to provide a tetravalent bispecific antibody. ADARPin can be appended to the C-termini of the light chains and theC-termini of the heavy chains of an IgG, to provide a hexavalentbispecific antibody. Additional examples of multispecific antibodiesproduced by cloning techniques include: (i) DVD-Ig™ (dual variabledomain immunoglobulin, tandem linkage of the second V_(H) and V_(L) tothe N-termini of HC and LC, respectively), (ii) Tandemab (tandem linkageof 2 V_(H)-C_(H)1 in combination of common LC), (iii) DNL (naturalassociation of 2 antibodies or antibody fragments anchored with DDD(dimerization and docking domain) from PKA (protein kinase A) and AD(anchoring domain) from A-kinase anchor protein (AKAP), respectively),(iv) LUZ-Y (leucine zipper tethered at the C-termini of HC and laterproteolytically removed), (v) 2-in-1-IgG (same LC and HC capable of dualrecognition), and (vi) mAb² (engineered loops in C_(H)3 domain of IgG toobtain second specificity).

Another class of multispecific antibodies can be characterized bystructures with variable domains or scFvs as the building blocks.Non-limiting examples of such multispecific antibodies include two V_(H)domains joined in tandem, diabodies (heterodimers containing 2polypeptide chains encoding V_(L)A-V_(H)B and V_(H)A-V_(L)B in the orderof V_(H)-V_(L) or V_(L)—V_(H) with a linker of 5 amino acids), dsDbs(interchain disulfide bond between V_(L) and V_(H) of the sameantibody), DARTs (dual-affinity re-targeting, interchain disulfide bondbetween 2 V_(L)), scDbs (single chain Diabody), tandAbs (Diabody dimervia flexible linkers in between), and 2 scFvs connected in tandem by anadjustable linker.

Another class of multispecific antibodies can contain different antigenbinding fragments, while retaining the basic IgG structure. Suchantibodies can comprise, for example, two distinct heavy chains and/ortwo distinct light chains. Various techniques can be used to promotepairing of desirable light and heavy chain combinations, rather thanrandom chain associations. Non-limiting examples of such techniquesinclude use of a common light chain, orthogonal Fab interface(complementary mutations introduced at LC and HC interface in one Faband no change to the other Fab), CrossMab (wherein one Fab V_(H) orC_(H)1 domain(s) can be switched with the partner V_(L) or C_(L)domain(s), with the other Fab untouched), and replacing the Fab with asingle chain antigen-binding domain. Further examples includeengineering strategies that can introduce mutations into the C_(H)3domains to promote heterodimerization based on steric or electrostaticcomplementarity. The “knobs in holes” approach can involve creating a“knob” by replacing threonine at position 366 with a bulky tryptophanresidue on one heavy chain, and making a corresponding “hole” by triplemutations (T366S, L368A and Y407V) on the partner heavy chain. Anotherapproach can involve creating alternating human IgG and IgA fragments inC_(H)3 to provide the so-called SEEDbody (Strand-Exchange EngineeredDomain) to guide heavy chain heterodimerization.

Non-limiting schematics of multispecific antibodies are provided inFIGS. 2-15 .

FIG. 2 provides a schematic of a tetravalent, bispecific antibody withsequences appended to the heavy chain C-termini. Light chain sequencesof the IgG isotype antibody unit are represented by “SEQ A”. Heavy chainsequences of the IgG isotype antibody unit are represented by “SEQ B”.Linker sequences are represented by “SEQ C”. Appended antigen bindingdomain sequences are represented by “SEQ D”. N and C denote N- andC-termini, respectively. —S—S— denotes disulfide bonds.

FIG. 3 provides a schematic of a tetravalent, bispecific antibody withsequences appended to the light chain C-termini. Light chain sequencesof the IgG isotype antibody unit are represented by “SEQ A”. Heavy chainsequences of the IgG isotype antibody unit are represented by “SEQ B”.Linker sequences are represented by “SEQ C”. Appended antigen bindingdomain sequences are represented by “SEQ D”. N and C denote N- andC-termini, respectively. —S—S— denotes disulfide bonds.

FIG. 4 provides a schematic of a hexavalent, bispecific antibody withsequences appended to the heavy chain and light chain C-termini. Lightchain sequences of the IgG isotype antibody unit are represented by “SEQA”. Heavy chain sequences of the IgG isotype antibody unit arerepresented by “SEQ B”. Linker sequences are represented by “SEQ C” and“SEQ E”. Appended antigen binding domain sequences are represented by“SEQ D”. N and C denote N- and C-termini, respectively. —S—S— denotesdisulfide bonds.

FIG. 5 provides a schematic of a hexavalent, trispecific antibody withsequences appended to the heavy chain and light chain C-termini. Lightchain sequences of the IgG isotype antibody unit are represented by “SEQA”. Heavy chain sequences of the IgG isotype antibody unit arerepresented by “SEQ B”. Linker sequences are represented by “SEQ C” and“SEQ E”. Appended antigen binding domain sequences are represented by“SEQ D” and “SEQ F”. N and C denote N- and C-termini, respectively.—S—S— denotes disulfide bonds.

FIG. 6 provides a schematic of a bivalent, bispecific antibody with twodifferent heavy chain sequences and two different light chain sequences.Light chain sequences are represented by “SEQ A” and “SEQ D”. Heavychain sequences are represented by “SEQ B” and “SEQ C”. N and C denoteN- and C-termini, respectively. —S—S— denotes disulfide bonds.

FIG. 7 provides a schematic of a trivalent, trispecific antibody withtwo different heavy chain sequences, two different light chainsequences, and a sequence appended to the C-terminus of one light chain.Light chain sequences of the IgG isotype antibody unit are representedby “SEQ A” and “SEQ D”. Heavy chain sequences of the IgG isotypeantibody unit are represented by “SEQ B” and “SEQ C”. A linker sequenceis represented by “SEQ E”. An appended antigen binding domain sequenceis represented by “SEQ F”. N and C denote N- and C-termini,respectively. —S—S— denotes disulfide bonds.

FIG. 8 provides a schematic of a trivalent, trispecific antibody withtwo different heavy chain sequences, two different light chainsequences, and a sequence appended to the C-terminus of one heavy chain.Light chain sequences of the IgG isotype antibody unit are representedby “SEQ A” and “SEQ D”. Heavy chain sequences of the IgG isotypeantibody unit are represented by “SEQ B” and “SEQ C”. A linker sequenceis represented by “SEQ E”. An appended antigen binding domain sequenceis represented by “SEQ F”. N and C denote N- and C-termini,respectively. —S—S— denotes disulfide bonds.

FIG. 9 provides a schematic of a tetravalent antibody with two differentheavy chain sequences, two different light chain sequences, andsequences appended to both light chain C-termini. Light chain sequencesof the IgG isotype antibody unit are represented by “SEQ A” and “SEQ D”.Heavy chain sequences of the IgG isotype antibody unit are representedby “SEQ B” and “SEQ C”. Linker sequences are represented by “SEQ E” and“SEQ F”. Appended antigen binding domain sequences are represented by“SEQ G” and “SEQ H”. N and C denote N- and C-termini, respectively.—S—S— denotes disulfide bonds.

FIG. 10 provides a schematic of a tetravalent antibody with twodifferent heavy chain sequences, two different light chain sequences,and sequences appended to both heavy chain C-termini. Light chainsequences of the IgG isotype antibody unit are represented by “SEQ A”and “SEQ D”. Heavy chain sequences of the IgG isotype antibody unit arerepresented by “SEQ B” and “SEQ C”. Linker sequences are represented by“SEQ E” and “SEQ F”. Appended antigen binding domain sequences arerepresented by “SEQ G” and “SEQ H”. N and C denote N- and C-termini,respectively. —S—S— denotes disulfide bonds.

FIG. 11 provides a schematic of a tetravalent antibody with twodifferent heavy chain sequences, two different light chain sequences,and sequences appended to one heavy chain C-terminus and one light chainC-terminus in cis. Light chain sequences of the IgG isotype antibodyunit are represented by “SEQ A” and “SEQ D”. Heavy chain sequences ofthe IgG isotype antibody unit are represented by “SEQ B” and “SEQ C”.Linker sequences are represented by “SEQ E” and “SEQ F”. Appendedantigen binding domain sequences are represented by “SEQ G” and “SEQ H”.N and C denote N- and C-termini, respectively. —S—S— denotes disulfidebonds.

FIG. 12 provides a schematic of a tetravalent antibody with twodifferent heavy chain sequences, two different light chain sequences,and sequences appended to one heavy chain C-terminus and one light chainC-terminus in trans. Light chain sequences of the IgG isotype antibodyunit are represented by “SEQ A” and “SEQ D”. Heavy chain sequences ofthe IgG isotype antibody unit are represented by “SEQ B” and “SEQ C”.Linker sequences are represented by “SEQ E” and “SEQ F”. Appendedantigen binding domain sequences are represented by “SEQ G” and “SEQ H”.N and C denote N- and C-termini, respectively. —S—S— denotes disulfidebonds.

FIG. 13 provides a schematic of a pentavalent antibody with twodifferent heavy chain sequences, two different light chain sequences,and sequences appended to both heavy chain C-termini and one light chainC-terminus. Light chain sequences of the IgG isotype antibody unit arerepresented by “SEQ A” and “SEQ D”. Heavy chain sequences of the IgGisotype antibody unit are represented by “SEQ B” and “SEQ C”. Linkersequences are represented by “SEQ E”, “SEQ F”, and “SEQ G”. Appendedantigen binding domain sequences are represented by “SEQ H”, “SEQ I”,and “SEQ J”. N and C denote N- and C-termini, respectively. —S—S—denotes disulfide bonds.

FIG. 14 provides a schematic of a pentavalent antibody with twodifferent heavy chain sequences, two different light chain sequences,and sequences appended to one heavy chain C-terminus and both lightchain C-termini. Light chain sequences of the IgG isotype antibody unitare represented by “SEQ A” and “SEQ D”. Heavy chain sequences of the IgGisotype antibody unit are represented by “SEQ B” and “SEQ C”. Linkersequences are represented by “SEQ E”, “SEQ F”, and “SEQ G”. Appendedantigen binding domain sequences are represented by “SEQ H”, “SEQ I”,and “SEQ J”. N and C denote N- and C-termini, respectively. —S—S—denotes disulfide bonds.

FIG. 15 provides a schematic of a hexavalent antibody with two differentheavy chain sequences, two different light chain sequences, andsequences appended to the heavy chain and light chain C-termini. Lightchain sequences of the IgG isotype antibody unit are represented by “SEQA” and “SEQ D”. Heavy chain sequences of the IgG isotype antibody unitare represented by “SEQ B” and “SEQ C”. Linker sequences are representedby “SEQ E”, “SEQ F”, “SEQ G”, and “SEQ H”. Appended antigen bindingdomain sequences are represented by “SEQ I”, “SEQ J”, “SEQ K”, and “SEQL”. N and C denote N- and C-termini, respectively. —S—S— denotesdisulfide bonds.

Antigen-binding compounds specific for, for example, HPTP-β (VE-PTP),can be combined with antigen-binding compounds specific for, forexample, a RTK agonist, to provide a polyvalent multispecific compoundthat can bind both HPTP-β (VE-PTP) and the RTK agonist. For example, anantigen binding compound specific for HPTP-β (VE-PTP) can be combinedwith an antigen-binding compound specific for VEGF, Ang1, Ang2, BDNF,EGF, FGF, HGF, IGF, insulin, MSP, NGF, NT-3, PDGF, or any combinationthereof, to provide a polyvalent multispecific compound that can bindboth HPTP-β (VE-PTP) and a RTK agonist.

Antigen-binding compounds specific for, for example, HPTP-β (VE-PTP),can be combined with antigen-binding compounds specific for, forexample, VEGF, to provide a polyvalent multispecific compound that canbind both HPTP-β (VE-PTP) and VEGF. Compounds that bind both HPTP-β(VE-PTP) and VEGF can inhibit HPTP-β (VE-PTP), activate Tie2, inhibitVEGF binding to VEGFRs, and inhibit VEGFR signaling.

Sequences derived from aflibercept, a recombinant protein comprising theVEGF-binding portions of human VEGF receptors 1 and 2, can be combinedwith antigen-binding compounds specific for HPTP-β (VE-PTP). Forexample, sequences derived from aflibercept can be fused to theC-termini of the heavy chains of a HPTP-β (VE-PTP)-specific antibody, toprovide a tetravalent, bispecific antibody (FIG. 2 ). Sequences derivedfrom aflibercept can be fused to the C-termini of the light chains of aHPTP-β (VE-PTP)-specific antibody, to provide a tetravalent, bispecificantibody (FIG. 3 ). Sequences derived from aflibercept can be fused tothe C-termini of the light chains and heavy chains of a HPTP-β(VE-PTP)-specific antibody, to provide a hexavalent, bispecific antibody(FIG. 4 ).

Sequences derived from brolucizumab, a humanized single-chain antibodyfragment (scFv) inhibitor of VEGF that binds to the receptor bindingsite of VEGF, can be combined with antigen-binding compounds specificfor HPTP-β (VE-PTP). For example, sequences from brolucizumab can befused to the C-termini of the heavy chains of a HPTP-β (VE-PTP)-specificantibody, to provide a tetravalent, bispecific antibody (FIG. 2 ).Sequences derived from brolucizumab can be fused to the C-termini of thelight chains of a HPTP-β (VE-PTP)-specific antibody, to provide atetravalent, bispecific antibody (FIG. 3 ). Sequences derived frombrolucizumab can be fused to the C-termini of the light chains and heavychains of a HPTP-β (VE-PTP)-specific antibody, to provide a hexavalent,bispecific antibody (FIG. 4 ).

Sequences derived from ranibizumab, a humanized monoclonal antibodyfragment (Fab) that binds to and inhibits the activity of VEGF, can becombined with antigen-binding compounds specific for HPTP-β (VE-PTP).For example, the sequences from ranibizumab can be cloned into an scFv,and the ranibizumab-derived scFv can be fused to can be fused to theC-termini of the heavy chains of a HPTP-β (VE-PTP)-specific antibody, toprovide a tetravalent, bispecific antibody (FIG. 2 ). Theranibizumab-derived scFv can be fused to the C-termini of the lightchains of a HPTP-β (VE-PTP)-specific antibody, to provide a tetravalent,bispecific antibody (FIG. 3 ). The ranibizumab-derived scFv can be fusedto the C-termini of the light chains and heavy chains of a HPTP-β(VE-PTP)-specific antibody, to provide a hexavalent, bispecific antibody(FIG. 4 ).

Sequences derived from bevacizumab, a humanized monoclonal antibody thatthat binds to and inhibits activity of VEGF, can be combined withantigen-binding compounds specific for HPTP-β (VE-PTP). For example, thesequences from bevacizumab can be cloned into an scFv, and thebevacizumab-derived scFv can be fused to can be fused to the C-terminiof the heavy chains of a HPTP-β (VE-PTP)-specific antibody, to provide atetravalent, bispecific antibody (FIG. 2 ). The bevacizumab-derived scFvcan be fused to the C-termini of the light chains of a HPTP-β(VE-PTP)-specific antibody, to provide a tetravalent, bispecificantibody (FIG. 3 ). The bevacizumab-derived scFv can be fused to theC-termini of the light chains and heavy chains of a HPTP-β(VE-PTP)-specific antibody, to provide a hexavalent, bispecific antibody(FIG. 4 ).

Sequences derived from conbercept, a recombinant protein comprising theVEGF-binding portions of VEGF receptors 1 and 2, can be combined withantigen-binding compounds specific for HPTP-β (VE-PTP). For example,sequences derived from conbercept can be fused to the C-termini of theheavy chains of a HPTP-β (VE-PTP)-specific antibody, to provide atetravalent, bispecific antibody (FIG. 2 ). Sequences derived fromconbercept can be fused to the C-termini of the light chains of a HPTP-β(VE-PTP)-specific antibody, to provide a tetravalent, bispecificantibody (FIG. 3 ). Sequences derived from conbercept can be fused tothe C-termini of the light chains and heavy chains of a HPTP-β(VE-PTP)-specific antibody, to provide a hexavalent, bispecific antibody(FIG. 4 ).

Sequences derived from abicipar, a VEGF-binding DARPin, can be combinedwith antigen-binding compounds specific for HPTP-β (VE-PTP). Forexample, sequences derived from abicipar can be fused to the C-terminiof the heavy chains of a HPTP-β (VE-PTP)-specific antibody, to provide atetravalent, bispecific antibody (FIG. 2 ). Sequences derived fromabicipar can be fused to the C-termini of the light chains of a HPTP-β(VE-PTP)-specific antibody, to provide a tetravalent, bispecificantibody (FIG. 3 ). Sequences derived from abicipar can be fused to theC-termini of the light chains and heavy chains of a HPTP-β(VE-PTP)-specific antibody, to provide a hexavalent, bispecific antibody(FIG. 4 ).

Antigen-binding compounds specific for HPTP-β (VE-PTP) can be combinedwith DARPins or amino acid sequences therefrom, for example, amino acidsequences comprising any one of SEQ ID NOS: 158-217. Amino acidsequences comprising any one of SEQ ID NOS: 158-217 can be fused to theC-termini of the heavy chains of a HPTP-β (VE-PTP)-specific antibody, toprovide a tetravalent, bispecific antibody (FIG. 2 ). Amino acidsequences comprising any one of SEQ ID NOS: 158-217 can be fused to theC-termini of the light chains of a HPTP-β (VE-PTP)-specific antibody, toprovide a tetravalent, bispecific antibody (FIG. 3 ). Amino acidsequences comprising any one of SEQ ID NOS: 158-217 can be fused to theC-termini of the light chains and heavy chains of a HPTP-β(VE-PTP)-specific antibody, to provide a hexavalent, bispecific antibody(FIG. 4 ).

Multiple VEGF-specific compounds can be combined with an antigen-bindingcompound specific for HPTP-β (VE-PTP). For example, one binding domain,(e.g. aflibercept-derived sequences) can be fused to the C-termini ofthe heavy chains of an anti-HPTP-β (VE-PTP) antibody, and anotherbinding domain (e.g. brolucizumab-derived sequences) be fused to theC-termini of the light chains of the HPTP-β (VE-PTP)-specific antibody,to provide a hexavalent, trispecific antibody (FIG. 5 ).

If different heavy chains are used in the basic four chain IgG antibodyunit (e.g., using the “knobs in holes” approach), antibodies can begenerated that are bivalent, trivalent, tetravalent, pentavalent, orhexavalent, and that are monospecific, bispecific, trispecific,tetraspecific, pentaspecific, or hexaspecific.

In one non-limiting example, one arm of the antibody unit can containCDRs specific for HPTP-β (VE-PTP), while the other arm of the antibodyunit can contain CDRs specific for VEGF (FIG. 6 ).

In one non-limiting example, both antigen binding fragments of the basicantibody unit can be HPTP-β (VE-PTP)-specific, and a binding domainspecific for VEGF (e.g. aflibercept-derived sequence,brolucizumab-derived sequence, ranibizumab-derived sequence, or abevacizumab-derived sequence) can be fused to one light chain, toprovide a trivalent bispecific antibody (FIG. 7 ).

In one non-limiting example, both antigen binding fragments of the basicantibody unit can be HPTP-β (VE-PTP)-specific, and a binding domainspecific for VEGF (e.g. aflibercept-derived sequence,brolucizumab-derived sequence, ranibizumab-derived sequence, or abevacizumab-derived sequence) can be fused to one heavy chain, toprovide a trivalent bispecific antibody (FIG. 8 ).

In one non-limiting example, both antigen binding fragments of the basicantibody unit can be HPTP-β (VE-PTP)-specific, and a binding domainspecific for VEGF (e.g. aflibercept-derived sequence,brolucizumab-derived sequence, ranibizumab-derived sequence, or abevacizumab-derived sequence) can be fused to one light chain, and asecond, different binding domain specific for VEGF can be fused to theother light chain, to provide a tetravalent trispecific antibody (FIG. 9).

In one non-limiting example, both antigen binding fragments of the basicantibody unit can be HPTP-β (VE-PTP)-specific, a binding domain specificfor VEGF (e.g. aflibercept-derived sequence, brolucizumab-derivedsequence, ranibizumab-derived sequence, or a bevacizumab-derivedsequence) can be fused to one heavy chain, and a second, differentbinding domain specific for VEGF can be fused to the other heavy chain,to provide a tetravalent trispecific antibody (FIG. 10 ).

In one non-limiting example, both antigen binding fragments of the basicantibody unit can be HPTP-β (VE-PTP)-specific, and a binding domainspecific for VEGF (e.g. aflibercept-derived sequence,brolucizumab-derived sequence, ranibizumab-derived sequence, or abevacizumab-derived sequence) can be fused to one heavy chain, and asecond, different binding domain specific for VEGF can be fused to onelight chain in cis, to provide a tetravalent trispecific antibody (FIG.11 ).

In one non-limiting example, both antigen binding fragments of the basicantibody unit can be HPTP-β (VE-PTP)-specific, a binding domain specificfor VEGF (e.g. aflibercept-derived sequence, brolucizumab-derivedsequence, ranibizumab-derived sequence, or a bevacizumab-derivedsequence) can be fused to one heavy chain, and a second, differentbinding domain specific for VEGF can be fused to one light chain intrans, to provide a tetravalent trispecific antibody (FIG. 12 ).

In one non-limiting example, both antigen binding fragments of the basicantibody unit can be HPTP-β (VE-PTP)-specific, a binding domain specificfor VEGF (e.g. aflibercept-derived sequence, brolucizumab-derivedsequence, ranibizumab-derived sequence, or a bevacizumab-derivedsequence) can be fused to one heavy chain, a second, different bindingdomain specific for VEGF can be fused to a second heavy chain, and athird, different binding domain specific for VEGF can be fused to onelight chain, to provide a pentavalent tetraspecific antibody (FIG. 13 ).

In one non-limiting example, both antigen binding fragments of the basicantibody unit can be HPTP-β (VE-PTP)-specific, a binding domain specificfor VEGF (e.g. aflibercept-derived sequence, brolucizumab-derivedsequence, ranibizumab-derived sequence, or a bevacizumab-derivedsequence) can be fused to one heavy chain, a second, different bindingdomain specific for VEGF can be fused to a one light chain, and a third,different binding domain specific for VEGF can be fused to the otherlight chain, to provide a pentavalent tetraspecific antibody (FIG. 14 ).

In one non-limiting example, both antigen binding fragments of the basicantibody unit can be HPTP-β (VE-PTP)-specific, a binding domain specificfor VEGF (e.g. aflibercept-derived sequence, brolucizumab-derivedsequence, ranibizumab-derived sequence, or a bevacizumab-derivedsequence) can be fused to one heavy chain, a second, different bindingdomain specific for VEGF can be fused to the other heavy chain, a third,different binding domain specific for VEGF can be fused to one lightchain, and a fourth binding domain specific for VEGF can be fused to theother light chain, to provide a hexavalent pentaspecific antibody (FIG.15 ).

In one non-limiting example, both antigen binding fragments of the basicantibody unit can be HPTP-β (VE-PTP)-specific, one binding domain, (e.g.an aflibercept-derived sequence) can be fused to the C-terminus of oneheavy chain, and second binding domain (e.g. a brolucizumab-derivedsequence) be fused to the C-terminus of the other heavy chain, toprovide a tetravalent, trispecific antibody (FIG. 10 ). In onenon-limiting example, both antigen binding fragments of the basicantibody unit can be HPTP-β (VE-PTP)-specific, an aflibercept-derivedsequence can be fused to the C-terminus of one light chain, and abrolucizumab-derived sequence be fused to the C-terminus of anotherlight chain, to provide a tetravalent, trispecific antibody (FIG. 9 ).In one non-limiting example, both antigen binding fragments of the basicantibody unit can be HPTP-β (VE-PTP)-specific, an aflibercept-derivedsequence can be fused to the C-terminus of one heavy chain, and abrolucizumab-derived sequence be fused to the C-terminus of one lightchain, to provide a tetravalent, trispecific antibody (FIG. 11 , FIG. 12). In one non-limiting example, both antigen binding fragments of thebasic antibody unit can be HPTP-β (VE-PTP)-specific, anaflibercept-derived sequence can be fused to the C-terminus of one heavychain, a brolucizumab-derived sequence can be fused to the C-terminus ofanother heavy chain, and ranibizumab-derived sequences can be fused tothe C-termini of both light chains, to provide a hexavalent,tetraspecific antibody (FIG. 15 ). In one non-limiting example, bothantigen binding fragments of the basic antibody unit can be HPTP-β(VE-PTP)-specific, an aflibercept-derived sequence can be fused to theC-terminus of one heavy chain, a brolucizumab-derived sequence can befused to the C-terminus of another heavy chain, a ranibizumab-derivedsequence can be fused to the C-terminus of one light chain, and abevacizumab-derived sequence can be fused to the C-terminus of anotherlight chain, to provide a hexavalent, pentaspecific antibody (FIG. 15 ).

Antigen-binding compounds specific for, for example, HPTP-β (VE-PTP),can be combined with other amino acid sequences to provide polyvalentmultispecific compounds that, for example, enhance Tie2 activation,enhance Tie2 phosphorylation, enhance Tie2 signaling, reduce VEGFRactivation, reduce VEGFR phosphorylation, reduce VEGFR signaling, or acombination thereof.

Antigen-binding compounds specific for HPTP-β (VE-PTP) can be combinedwith collagen IV-derived biomimetic peptides, for example, amino acidsequences comprising SEQ ID NO: 152 or SEQ ID NO: 153. Amino acidsequences comprising SEQ ID NO: 152 or SEQ ID NO: 153 can be fused tothe C-termini of the heavy chains of a HPTP-β (VE-PTP)-specificantibody, to provide a tetravalent, bispecific antibody (FIG. 2 ). Aminoacid sequences comprising SEQ ID NO: 152 or SEQ ID NO: 153 can be fusedto the C-termini of the light chains of a HPTP-β (VE-PTP)-specificantibody, to provide a tetravalent, bispecific antibody (FIG. 3 ). Aminoacid sequences comprising SEQ ID NO: 152 or SEQ ID NO: 153 the C-terminiof the light chains and heavy chains of a HPTP-β (VE-PTP)-specificantibody, to provide a hexavalent, bispecific antibody (FIG. 4 ).

Antigen-binding compounds specific for HPTP-β (VE-PTP) can be combinedwith Ang1 mimetics, for example, vasculotide. Sequences derived fromvasculotide can be fused to the C-termini of the heavy chains of aHPTP-β (VE-PTP)-specific antibody, to provide a tetravalent, bispecificantibody (FIG. 2 ). Sequences derived from vasculotide can be fused tothe C-termini of the light chains of a HPTP-β (VE-PTP)-specificantibody, to provide a tetravalent, bispecific antibody (FIG. 3 ).Sequences derived from vasculotide can be fused to the C-termini of thelight chains and heavy chains of a HPTP-β (VE-PTP)-specific antibody, toprovide a hexavalent, bispecific antibody (FIG. 4 ).

Antigen-binding compounds specific for, for example, HPTP-β (VE-PTP),can be combined with antigen-binding compounds specific for, forexample, a RTK, to provide a polyvalent multispecific compound that canbind both HPTP-β (VE-PTP) and the RTK. For example, an antigen bindingcompound specific for HPTP-β (VE-PTP) can be combined with anantigen-binding compound specific for VEGFR (e.g. VEGFR2), to provide apolyvalent multispecific compound that can bind both HPTP-β (VE-PTP) andVEGFR.

Compounds that bind both HPTP-β (VE-PTP) and VEGFR can inhibit HPTP-β(VE-PTP), activate Tie2, inhibit VEGF binding to VEGFR, and inhibitVEGFR signaling.

Sequences derived from ramucirumab, a humanized monoclonal antibody thatbinds an extracellular domain of VEGFR2 and inhibits VEGFR2 signaling,can be combined with antigen-binding compounds specific for HPTP-β(VE-PTP). For example, the sequences from ramucirumab can be cloned intoan scFv, and the ramucirumab-derived scFv can be fused to can be fusedto the C-termini of the heavy chains of a HPTP-β (VE-PTP)-specificantibody, to provide a tetravalent, bispecific antibody (FIG. 2 ). Theramucirumab-derived scFv can be fused to the C-termini of the lightchains of a HPTP-β (VE-PTP)-specific antibody, to provide a tetravalent,bispecific antibody (FIG. 3 ). The ramucirumab-derived scFv can be fusedto the C-termini of the light chains and heavy chains of a HPTP-β(VE-PTP)-specific antibody, to provide a hexavalent, bispecific antibody(FIG. 4 ).

Multiple compounds that enhance Tie2 activation, enhance Tie2phosphorylation, enhance Tie2 signaling, reduce VEGFR activation, reduceVEGFR phosphorylation, reduce VEGFR signaling, or a combination thereof,can be combined with an antigen-binding compound specific for HPTP-β(VE-PTP). For example, one binding domain, (e.g. brolucizumab-derivedsequences) can be fused to the C-termini of the heavy chains of ananti-HPTP-β (VE-PTP) antibody, and another binding domain (e.g.vasculotide-derived sequences) be fused to the C-termini of the lightchains of the HPTP-β (VE-PTP)-specific antibody, to provide ahexavalent, trispecific antibody (FIG. 5 ).

Compounds, antibodies, or derivatives thereof disclosed herein can have1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 domains or more. Each domaincan modulate, bind, antagonize, inhibit or activate any targetsdisclosed herein, for example, a phosphatase, a phosphatase thatmodulates Tie2 signaling, a protein tyrosine phosphatase, areceptor-like protein tyrosine phosphatase, a Tie2 modulator, HPTP-β(VE-PTP), an extracellular domain of HPTP-β (VE-PTP), the first FN3repeat of an extracellular domain of HPTP-β (VE-PTP), a kinase, atyrosine kinase, a receptor tyrosine kinase, a receptor tyrosine kinaseactivator, a receptor tyrosine kinase agonist, a growth factor, a growthfactor receptor activator, a growth factor receptor agonist, acysteine-knot growth factor superfamily member, a pro-angiogenic factor,a PDGF family member, a VEGF receptor, a VEGF receptor agonist, a VEGFfamily member, a VEGF, VEGF-A, VEGF-B, VEGF-C, VEGF-D, PGF, VEGF₁₂₁,VEGF₁₄₅, VEGF₁₄₈, VEGF₁₆₂, VEGF₁₆₅, VEGF_(165b), VEGF₁₈₃, VEGF₁₈₉ orVEGF₂₀₆.

Inhibitors, Activators, Modulators, and Binding Agents

A HPTP-β (VE-PTP) inhibitor, modulator, or binding agent of thedisclosure can include a compound, a recombinant protein, an antibody,antigen-binding fragment, variant, or derivative thereof, aTie2-peptomimetic, a tetrameric polyethylene oxide clustered peptide, acollagen IV-biomimetic peptide, a DARPin or derivative thereof, anaffinibody, an avimer, an adnectin, an anticalin, a Fynomer, a Kunitzdomain, a knottin, a β-hairpin mimetic, or a peptide derived from one ormore receptors, either alone or in combination with another amino acidsequence or multiple other amino acid sequences. The inhibitor,modulator, or binding agent can undergo modifications, for example,enzymatic cleavage or posttranslational modifications.

In some embodiments, a HPTP-β (VE-PTP) inhibitor, modulator, or bindingagent of the disclosure can inhibit HPTP-β (VE-PTP) by interfering withthe interaction of HPTP-β (VE-PTP) and Tie2. In some embodiments, aHPTP-β (VE-PTP) inhibitor, modulator, or binding agent of the disclosurecan inhibit HPTP-β (VE-PTP) by stabilizing HPTP-β (VE-PTP) in aninactive conformation. In some embodiments, a HPTP-β (VE-PTP) inhibitor,modulator, or binding agent of the disclosure can inhibit HPTP-β(VE-PTP) by promoting internalization of HPTP-β (VE-PTP) (e.g.,promoting endocytosis and degradation of HPTP-β (VE-PTP)). In someembodiments, a HPTP-β (VE-PTP) inhibitor, modulator, or binding agent ofthe disclosure can inhibit HPTP-β (VE-PTP) by blocking binding of aligand that activates HPTP-β (VE-PTP). In some embodiments, a HPTP-β(VE-PTP) inhibitor, modulator, or binding agent of the disclosure caninhibit HPTP-β (VE-PTP) by modulating oligomerization of HPTP-β(VE-PTP).

In some embodiments, a HPTP-β (VE-PTP) inhibitor, modulator, or bindingagent of the disclosure can bind to a dominant-negative isoform ofHPTP-β (VE-PTP). A dominant-negative isoform can correspond to a form ofHPTP-β (VE-PTP) deficient in phosphatase activity that can compete withendogenous HPTP-β (VE-PTP).

A HPTP-β (VE-PTP) inhibitor, modulator, or binding agent of thedisclosure can comprise a plurality of HPTP-β (VE-PTP) binding sites. Insome embodiments, a HPTP-β (VE-PTP) inhibitor, modulator, or bindingagent can bind to two HPTP-β (VE-PTP) molecules simultaneously, therebybringing the two HPTP-β (VE-PTP) molecules into close proximity. In someembodiments, a HPTP-β (VE-PTP) inhibitor, modulator, or binding agentcan bind to three HPTP-β (VE-PTP) molecules simultaneously, therebybringing the three HPTP-β (VE-PTP) molecules into close proximity. Insome embodiments, a HPTP-β (VE-PTP) inhibitor, modulator, or bindingagent can bind to four HPTP-β (VE-PTP) molecules simultaneously, therebybringing the four HPTP-β (VE-PTP) molecules into close proximity.

A HPTP-β (VE-PTP) inhibitor, modulator, or binding agent of thedisclosure can be covalently or non-covalently conjugated to anothermoiety or vehicle. A moiety or vehicle can, for example, provide bindingspecificity for an additional target, inhibit degradation, increasehalf-life, increase absorption, reduce toxicity, reduce immunogenicity,and/or increase biological activity of the inhibitor, modulator, orbinding agent. Non-limiting examples of the moiety to which theinhibitor, modulator, or binding agent can be conjugated include a Fcdomain of an immunoglobulin, a peptide, a lipid, a carbohydrate, adendrimer, an oligosaccharide, a cholesterol group such as a steroid,and a polymer such as a polyethylene glycol (PEG), a polylysine, or adextran.

A compound of the present disclosure can be used for targeting HPTP-β(VE-PTP) to restore Tie2 activity. A HPTP-β (VE-PTP) inhibitor,modulator, or binding agent can thus be a Tie2 activator. In someembodiments, a compound of the present disclosure can initiate orinhibit a signaling cascade downstream of HPTP-β (VE-PTP) or Tie2, forexample, Akt/PI3-K signaling, Rac1 signaling, MAPK/Ras signaling, orNF-κB signaling.

Inhibition of HPTP-β (VE-PTP) can lead to vascular stabilization, whichcan be beneficial for the treatment of, for example, disorders that arecharacterized by vascular instability, angiogenesis, neovascularization,vascular leakage, and/or edema. For example, inhibition of HPTP-β(VE-PTP) can be beneficial for the treatment of vascular disorders,ocular disorders, cancers, renal disorders, complications of diabetes,and other disorders. In some embodiments, a HPTP-β (VE-PTP) inhibitor,modulator, or binding agent of the disclosure can be used to treat, forexample, diabetic retinopathy, non-proliferative diabetic retinopathy(NPDR), glaucoma, intraocular pressure, ocular edema, ocular hemorrhage,ocular hypertension, ocular inflammation, ocular neovascularization,ocular vascular leak, retinal perfusion, or retinopathy.

A Tie2 activator, modulator, or binding agent of the disclosure caninclude, for example, a compound, a recombinant protein, a peptide, anantibody, an antigen-binding fragment, variant, or derivative thereof,an angiopoietin 1 recombinant protein, an Ang1 mimetic, a Tie2 agonist,a HPTP-β (VE-PTP) phosphatase inhibitor, a Tie2-peptomimetic, atetrameric polyethylene oxide clustered peptide, a collagenIV-biomimetic peptide, a DARPin or derivative thereof, an affinibody, anavimer, an adnectin, an anticalin, a Fynomer, a Kunitz domain, aknottin, a β-hairpin mimetic, or a peptide derived from one or morereceptors, either alone or in combination with another amino acidsequence or multiple other amino acid sequences. The activator,modulator, or binding agent can undergo modifications, for example,enzymatic cleavage or posttranslational modifications.

A Tie2 activator, modulator, or binding agent of the disclosure can becovalently or non-covalently conjugated to another moiety or vehicle. Amoiety or vehicle can, for example, provide binding specificity for anadditional target, inhibit degradation, increase half-life, increaseabsorption, reduce toxicity, reduce immunogenicity, and/or increasebiological activity of the activator, modulator, or binding agent.Non-limiting examples of the moiety to which the activator, modulator,or binding agent can be conjugated include a Fc domain of animmunoglobulin, a peptide, a lipid, a carbohydrate, a dendrimer, anoligosaccharide, a cholesterol group such as a steroid, and a polymersuch as a polyethylene glycol (PEG), a polylysine, or a dextran.

In some embodiments, a compound of the present disclosure can initiateor inhibit a signaling cascade downstream of Tie2, for example,Akt/PI3-K signaling, Rac1 signaling, MAPK/Ras signaling, or NF-κBsignaling.

The activation of Tie2 can lead to vascular stabilization, which can bebeneficial for the treatment of, for example, disorders that arecharacterized by vascular instability, angiogenesis, neovascularization,vascular leakage, and/or edema. For example, activation of Tie2 can bebeneficial for the treatment of ocular disorders, cancers, renaldisorders, complications of diabetes, and other disorders. In someembodiments, a Tie2 activator, modulator, or binding agent of thedisclosure can be used to treat diabetic retinopathy, non-proliferativediabetic retinopathy (NPDR), glaucoma, intraocular pressure, ocularedema, ocular hemorrhage, ocular hypertension, ocular inflammation,ocular neovascularization, ocular vascular leak, retinal perfusion, orretinopathy.

A VEGF inhibitor, modulator, or binding agent of the disclosure caninclude a compound, a recombinant protein, a peptide, an antibody,antigen-binding fragment, variant, or derivative thereof, a DARPin orderivative thereof, an affinibody, an avimer, an adnectin, an anticalin,a Fynomer, a Kunitz domain, a knottin, a β-hairpin mimetic, a tetramericpolyethylene oxide clustered peptide, a collagen IV-biomimetic peptide,or a peptide derived from one or more receptors (e.g. VEGF receptors, orthe VEGF-binding portions of human VEGF receptors 1 and 2), either aloneor in combination with another amino acid sequence or multiple otheramino acid sequences. The inhibitor, modulator, or binding agent canundergo modifications, for example, enzymatic cleavage orposttranslational modifications.

A VEGF inhibitor, modulator, or binding agent of the disclosure cancomprise a plurality of VEGF binding sites. In some embodiments, a VEGFinhibitor, modulator, or binding agent can bind to two VEGF moleculessimultaneously, thereby bringing the two VEGF molecules into closeproximity. In some embodiments, a VEGF inhibitor, modulator, or bindingagent can bind to three VEGF molecules simultaneously, thereby bringingthe three VEGF molecules into close proximity. In some embodiments, aVEGF inhibitor, modulator, or binding agent can bind to four VEGFmolecules simultaneously, thereby bringing the four VEGF molecules intoclose proximity.

A VEGF inhibitor, modulator, or binding agent of the disclosure can becovalently or non-covalently conjugated to another moiety or vehicle. Amoiety or vehicle can, for example, provide binding specificity for anadditional target, inhibit degradation, increase half-life, increaseabsorption, reduce toxicity, reduce immunogenicity, and/or increasebiological activity of the inhibitor, modulator, or binding agent.Non-limiting examples of the moiety to which the inhibitor, modulator,or binding agent can be conjugated include a Fc domain of animmunoglobulin, a peptide, a lipid, a carbohydrate, a dendrimer, anoligosaccharide, a cholesterol group such as a steroid, and a polymersuch as a polyethylene glycol (PEG), a polylysine, or a dextran.

A VEGFR inhibitor, modulator, or binding agent of the disclosure caninclude a compound, a recombinant protein, an antibody, anantigen-binding fragment, variant, or derivative thereof, a tetramericpolyethylene oxide clustered peptide, a collagen IV-biomimetic peptide,a DARPin or derivative thereof, an affinibody, an avimer, an adnectin,an anticalin, a Fynomer, a Kunitz domain, a knottin, a β-hairpinmimetic, or a peptide derived from one or more receptors, either aloneor in combination with another amino acid sequence or multiple otheramino acid sequences. The inhibitor, modulator, or binding agent canundergo modifications, for example, enzymatic cleavage orposttranslational modifications.

A VEGFR inhibitor, modulator, or binding agent of the disclosure cancomprise a plurality of VEGFR binding sites. In some embodiments, aVEGFR inhibitor, modulator, or binding agent can bind to two VEGFRmolecules simultaneously, thereby bringing the two VEGFR molecules intoclose proximity. In some embodiments, a VEGFR inhibitor, modulator, orbinding agent can bind to three VEGFR molecules simultaneously, therebybringing the three VEGFR molecules into close proximity. In someembodiments, a VEGFR inhibitor, modulator, or binding agent can bind tofour VEGFR molecules simultaneously, thereby bringing the four VEGFRmolecules into close proximity.

A VEGFR inhibitor, modulator, or binding agent of the disclosure can becovalently or non-covalently conjugated to another moiety or vehicle. Amoiety or vehicle can, for example, provide binding specificity for anadditional target, inhibit degradation, increase half-life, increaseabsorption, reduce toxicity, reduce immunogenicity, and/or increasebiological activity of the inhibitor, modulator, or binding agent.Non-limiting examples of the moiety to which the inhibitor, modulator,or binding agent can be conjugated include a Fc domain of animmunoglobulin, a peptide, a lipid, a carbohydrate, a dendrimer, anoligosaccharide, a cholesterol group such as a steroid, and a polymersuch as a polyethylene glycol (PEG), a polylysine, or a dextran.

In some embodiments, a VEGFR inhibitor, modulator, or binding agent ofthe disclosure can inhibit VEGFR by stabilizing VEGFR in an inactiveconformation. In some embodiments, a VEGFR inhibitor, modulator, orbinding agent of the disclosure can inhibit VEGFR by promotinginternalization of VEGFR (e.g., promoting endocytosis and degradation ofVEGFR). In some embodiments, a VEGFR inhibitor, modulator, or bindingagent of the disclosure can inhibit VEGFR by blocking binding of aligand that activates VEGFR (e.g., blocking VEGF ligation of VEGFR). Insome embodiments, a VEGFR inhibitor, modulator, or binding agent of thedisclosure can inhibit VEGFR by modulating oligomerization of VEGFR(e.g., preventing or reducing the likelihood of dimerization oroligomerization of VEGFR).

A compound of the present disclosure can be used for interfering withthe interaction of VEGF and VEGFR, thereby reducing VEGFRphosphorylation and downstream signaling. In some embodiments,inhibition of VEGF can reduce aberrant vasculogenesis, angiogenesis, orblood vessel permeabilization, thereby reducing pathologic vascularinstability. In some embodiments, inhibition of VEGF can be beneficialfor the treatment of disorders that are characterized by vascularinstability, angiogenesis, neovascularization, vascular leakage, and/oredema. For example, inhibition of VEGF can be beneficial for thetreatment of vascular disorders, ocular disorders, cancers, renaldisorders, complications of diabetes, and other disorders. In someembodiments, a VEGF inhibitor, modulator, or binding agent of thedisclosure can be used to treat diabetic retinopathy, non-proliferativediabetic retinopathy (NPDR), glaucoma, intraocular pressure, ocularedema, ocular hemorrhage, ocular hypertension, ocular inflammation,ocular neovascularization, ocular vascular leak, retinal perfusion, orretinopathy. In some embodiments, inhibition of VEGF can reduce cancer.

Methods

The promotion of Tie2-signaling and inhibition of VEGFR signaling canlead to vascular stabilization, which can be beneficial for thetreatment of a condition with a vascular component. A compound disclosedherein can be used to treat, for example, a disease characterized bychanges in the vasculature, a disease characterized by decreased Tie2activation, or a disease characterized by involvement of VEGF inpathogenesis, whether progressive or non-progressive, acute or chronic.

In some embodiments, a compound disclosed herein can be used to treat anocular disorder. A compound disclosed herein can be used to treat, forexample, age-related macular degeneration (dry form), age-relatedmacular degeneration (wet form), atopic keratitis, Bests disease,blepharitis, blurry vision, choroidal neovascularization, chronicretinal detachment, chronic uveitis/vitritis, choroiditis,conjunctivitis, contact lens overwear, corneal graft neovascularization,corneal graft rejection, corneal neovascularization, cystoid macularedema, diabetic macular edema, double vision, diabetic retinopathy,diseases associated with rubeosis (neovascularization of the angle),diseases caused by abnormal proliferation of fibrovascular or fibroustissue including all forms of proliferative vitreoretinopathy, Ealesdisease, elevated intraocular pressure, epidemic keratoconjunctivitis,floaters, glaucoma, hard yellow exudates within 500 m of the center ofthe fovea with adjacent retinal thickening, hyperviscosity syndromes,infections causing choroiditis, infections causing retinitis, irisneovascularization, ischemic retinopathy, loss of contrast, maculartelangectasia, mariginal keratolysis, multifocal choroiditis, myopia,neovascular glaucoma, non-proliferative diabetic retinopathy (NPDR),ocular edema, ocular hemorrhage, ocular histoplasmosis, ocularhypertension, ocular inflammation, ocular ischemia, ocularneovascularization, ocular trauma, ocular vascular leak, optic pits,papilloedema, pars planitis, phylectenulosis, polypoidal choroidalvasculopathy, post-laser complications, proliferative diabeticretinopathy, pterygium keratitis sicca, radial keratotomy, retinalangiomatous proliferation, retinal degeneration, retinal edema(including macular edema), retinal neovascularization, retinalperfusion, retinal thickening within 1 disc diameter of the center ofthe fovea, retinal thickening within 500 m of the center of the fovea,retinal vein occlusion (central or branch), retinitis, retinitispigmentosa, retinopathy, retinopathy of prematurity, scleritis,Stargarts disease, superior limbic keratitis, surgery induced edema,surgery induced neovascularization, Terrien's marginal degeneration,trachoma, trauma, uveitis, vasculitis (e.g. central retinal veinocclusion), or other ophthalmic diseases wherein the eye disease ordisorder is associated with ocular neovascularization, vascular leakage,or retinal edema, or a combination thereof.

In some embodiments, a compound disclosed herein can be used to treat acomplication of diabetes (e.g., a comorbidity of diabetes). A compounddisclosed herein can be used to treat, for example, Acuteglomerulonephritis, Acute myocardial infarction, Amputation, Amyotrophy,Aneurysm, Angina pectoris, Aortic aneurysm, Aortic dissection,Atherosclerosis, Atherosclerotic cardiovascular disease, Atrialfibrillation, Autonomic neuropathy, Blindness, Cardiovascularcomplications of diabetes, Cerebrovascular complications of diabetes,Charcot's arthropathy, Chronic glomerulonephritis, Chronic renalfailure, Claudication, Clinically significant macular edema, Coronaryartery disease, Cranial nerve palsy, Cystoid macular degeneration,Cystoid macular edema, Diabetic cardiomyopathy, Diabeticcheiroarthropathy, Diabetic coma, Diabetic encephalopathy, Diabetic footwound, Diabetic hyperglycemia, Diabetic hyperlipidemia, Diabetichyperosmolar syndrome, Diabetic hypoglycemia, Diabetic ketoacidosis,Diabetic myonecrosis, Diabetic nephropathy, Diabetic neuropathy,Diabetic ophthalmologic disease, Diabetic peripheral vascular disease,Diabetic retinopathy, Diffuse idiopathic skeletal hyperostosis,Dupuytren's contracture, Embolism, End-stage renal disease, Erectiledysfunction, Forestier disease, Gangrene, Gas gangrene,Gastroparesis/diarrhea, Heart failure, Hyperglycemic crisis,Hypertension, Ischemic heart disease, Ketoacidosis, Lipohypertrophy,Metabolic complications of diabetes, Mononeuropathy, Myocardialinfarction, Nephritis, Nephropathy, Nephrosis, Nephrotic syndrome,Neurogenic bladder, Neuropathic arthropathy, Neuropathy, Orthostatichypotension, Osteoarthritis, Osteoporosis, Periodontal disease,Peripheral vascular disease, Polyneuropathy, Proliferative retinopathy,Renal failure, Renal insufficiency, Restrictive lung disease, Retinaldetachment, Retinal edema, Retinopathy, Stroke, Thrombosis, Transientischemic attack, Ulceration, Ventricular fibrillation, Vitreoushemorrhage, or a combination thereof.

In some embodiments, a compound disclosed herein can be used to treat arenal disorder. A compound disclosed herein can be used to treat, forexample, Acute kidney injury, Acute proliferative glomerulonephritis,Adenine phosphoribosyltransferase deficiency, Alport syndrome, Analgesicnephropathy, Autosomal dominant polycystic kidney disease, Autosomalrecessive polycystic kidney disease, Balkan endemic nephropathy, Benignnephrosclerosis, Bright's disease, Cardiorenal syndrome, CFHR5nephropathy, Chronic kidney disease, Chronic kidney disease-mineral andbone disorder, Congenital nephrotic syndrome, Conorenal syndrome,Contrast-induced nephropathy, Cystic kidney disease, Dents disease,Diabetic nephropathy, Diffuse proliferative nephritis, Distal renaltubular acidosis, Diuresis, EAST syndrome, End Stage Renal Disease,Epithelial-mesenchymal transition, Epstein syndrome, Fanconi syndrome,Fechtner syndrome, Focal proliferative nephritis, Focal segmentalglomerulosclerosis, Fraley syndrome, Galloway Mowat syndrome, Gitelmansyndrome, Glomerulocystic kidney disease, Glomerulopathy, Goodpasturesyndrome, High anion gap metabolic acidosis, HIV-associated nephropathy,Horseshoe kidney, Hydronephrosis, Hypertensive kidney disease, IgAnephropathy, Interstitial nephritis, Juvenile nephronophthisis, Kidneycancer, Kidney disease, Kidney stone disease, Lightwood-Albrightsyndrome, Lupus nephritis, Malarial nephropathy, Medullary cystic kidneydisease, Medullary sponge kidney, Membranous glomerulonephritis,Mesoamerican nephropathy, Milk-alkali syndrome, Minimal mesangialglomerulonephritis, Multicystic dysplastic kidney, Nephritis,Nephrocalcinosis, Nephrogenic diabetes insipidus, Nephromegaly,Nephroptosis, Nephrosis, Nephrotic syndrome, Nutcracker syndrome,Papillorenal syndrome, Phosphate nephropathy, Polycystic kidney disease,Primary hyperoxaluria, Proximal renal tubular acidosis, Pyelonephritis,Pyonephrosis, Rapidly progressive glomerulonephritis, Renal agenesis,Renal angina, Renal artery stenosis, Renal cyst, Renal ischemia, Renalosteodystrophy, Renal papillary necrosis, Renal tubular acidosis, Renalvein thrombosis, Secondary hypertension, Serpentine fibula-polycystickidney syndrome, Shunt nephritis, Sickle cell nephropathy, Thin basementmembrane disease, Transplant glomerulopathy, Tubulointerstitialnephritis and uveitis, Tubulopathy, Uremia, Uremic frost, Wunderlichsyndrome, or a combination thereof.

In some embodiments, a compound disclosed herein can be used to treat acancer. A compound disclosed herein can be used to treat, for example,acute leukemia, astrocytomas, biliary cancer (cholangiocarcinoma), bonecancer, breast cancer, brain stem glioma, bronchioloalveolar cell lungcancer, cancer of the adrenal gland, cancer of the anal region, cancerof the bladder, cancer of the endocrine system, cancer of the esophagus,cancer of the head or neck, cancer of the kidney, cancer of theparathyroid gland, cancer of the penis, cancer of the pleural/peritonealmembranes, cancer of the salivary gland, cancer of the small intestine,cancer of the thyroid gland, cancer of the ureter, cancer of theurethra, carcinoma of the cervix, carcinoma of the endometrium,carcinoma of the fallopian tubes, carcinoma of the renal pelvis,carcinoma of the vagina, carcinoma of the vulva, cervical cancer,chronic leukemia, colon cancer, colorectal cancer, cutaneous melanoma,ependymoma, epidermoid tumors, Ewings sarcoma, gastric cancer,glioblastoma, glioblastoma multiforme, glioma, hematologic malignancies,hepatocellular (liver) carcinoma, hepatoma, Hodgkin's Disease,intraocular melanoma, Kaposi sarcoma, lung cancer, lymphomas,medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma,muscle cancer, neoplasms of the central nervous system (CNS), neuronalcancer, non-small cell lung cancer, osteosarcoma, ovarian cancer,pancreatic cancer, pediatric malignancies, pituitary adenoma, prostatecancer, rectal cancer, renal cell carcinoma, sarcoma of soft tissue,schwanoma, skin cancer, spinal axis tumors, squamous cell carcinomas,stomach cancer, synovial sarcoma, testicular cancer, uterine cancer, ortumors and their metastases, including refractory versions of any of theabove cancers, or a combination thereof.

In some embodiments, a compound disclosed herein can be used to treat adisease characterized by changes in the vasculature, a diseasecharacterized by decreased Tie2 activation, or a disease characterizedby involvement of VEGF in pathogenesis. A compound disclosed herein canbe used to treat, for example, acne rosacea, acute lung injury, acuterespiratory distress syndrome (ARDS), adhesion formation from abdominalsurgery, adipositas, albuminuria, allergic edema, allergy, angina,angiofibroma, arteriosclerosis, artery occlusion, ascites, atheroma,atherosclerosis, asthma, avascular necrosis, bacterial ulcers,Bartonella bacilliformis infection, Behcet's disease, Buerger's disease(thromboangiitis obliterans), cardiac fibrosis, cardiac hypertrophy,cardiomyopathy, carotid obstructive disease, cerebral infarction,chemical burns, COPD, Crohn's disease, cytokine-induced vascular leak,destabilized blood flow, diabetes (including non-insulin dependentdiabetes mellitus), dysfunctional uterine bleeding, endometriosis,Epstein-Barr virus infection, erectile dysfunction, excessive hairgrowth, follicular cysts, foot ulcer (e.g., diabetic foot ulcer), fungalulcers, giant cell arteritis, glomerulosclerosis, Graves' disease,Hashimoto's autoimmune thyroiditis, hemangioma, hemangioendothelioma,hemophilic joints, hemorrhage, hepatitis C, hereditary hemorrhagictelangiectasia (HHT), Herpes simplex infections, Herpes zosterinfections, hypertension, idiopathic thrombocytopenic purpura, impairedwound healing, inflammatory and infectious processes (e.g. hepatitis,pneumonia, glomerulonephritis), interstitial fibrosis, ischemia, kidneydisease, leishmaniasis, leukomalacia, lipid degeneration, liverregeneration, Lupus nephritis, Lyme disease, lymphoproliferativedisorders, malaria (Plasmodium infection), Mooren ulcer, multiplesclerosis, mycobacterial infections, myocardial infarction, nasalpolyps, nephropathy, neuronal inflammation, neuropathy, obesity,osteomyelitis, osteophyte, ovarian hyperstimulation, Paget's disease,pannus growth, peripheral artery disease, peritoneal sclerosis,pemphigoid, polyarteritis, protozoan infections, pseudoxanthomaelasticum, psoriasis, pulmonary hypertension, pyogenic granulomas, renalfibrosis, respiratory distress, rheumatoid arthritis, rickettsialinfection, scar keloids, sepsis, sickle cell anemia, Stevens-Johnsondisease, stroke, synovitis, systemic lupus erythematosus, syphilis,thyroid enlargement, thyroiditis, toxic shock syndrome, toxoplasmosis,trauma, ulcerative colitis, vascular leak, vascular leak syndrome,vascular malformations (e.g. Osler-Weber syndrome), vein occlusion,viral hemorrhagic fevers (e.g., dengue fever), vitamin A deficiency,warts, or Wegener's sarcoidosis, or a combination thereof.

Sequences

As used herein, the abbreviations for the L-enantiomeric andD-enantiomeric amino acids are as follows: alanine (A, Ala); arginine(R, Arg); asparagine (N, Asn); aspartic acid (D, Asp); cysteine (C,Cys); glutamic acid (E, Glu); glutamine (Q, Gln); glycine (G, Gly);histidine (H, His); isoleucine (I, Ile); leucine (L, Leu); lysine (K,Lys); methionine (M, Met); phenylalanine (F, Phe); proline (P, Pro);serine (S, Ser); threonine (T, Thr); tryptophan (W, Trp); tyrosine (Y,Tyr); valine (V, Val). In some embodiments, the amino acid is aL-enantiomer. In some embodiments, the amino acid is a D-enantiomer.

TABLE 2 shows the amino acid sequences of humanized V_(H) antibodyregions that bind HPTP-β (VE-PTP). SEQ ID NO: 1 is V_(H)1, SEQ ID NO: 2is V_(H)2, SEQ ID NO: 3 is V_(H)3, and SE ID NO: 4 is V_(H4).

TABLE 2 SEQ ID NO: Name Amino acid sequence 1 V_(H1)EVQLVESGGGLVQPGGSLKLSCAASGFTFNA NAMNWVRQASGKGLEWVGRIRTKSNNYATYYAGSVKDRFTISRDDSKNTAYLQMNSLKTEDT AAYYCVRDYYGSSAWITYWGQGTLVTVSS 2 V_(H2)EVQLVESGGGLVQPGGSLRLSCAASGFTFNA NAMNWVRQAPGKGLEWVGRIRTKSNNYATYYAGSVKDRFTISRDDSKNSLYLQMNSLKTEDT AVYYCVRDYYGSSAWITYWGQGTLVTVSS 3 V_(H3)EVQLVESGGGLVQPGRSLRLSCTASGFTFNA NAMNWVRQAPGKGLEWVGRIRTKSNNYATYYAGSVKDRFTISRDDSKNIAYLQMNSLKTEDT AVYYCVRDYYGSSAWITYWGQGTLVTVSS 4 V_(H4)LVQLVESGGGLVKPGGSLRLSCAASGFTFNA NAMNWIRQAPGKGLEWVSRIRTKSNNYATYYAGSVKDRFTISRDNAKNSLYLQMNSLRAEDT AVHYCVRDYYGSSAWITYWGQGTLVTVSS

TABLE 3 shows the amino acid sequences of humanized V_(L) antibodyregions that bind HPTP-β (VE-PTP). SEQ ID NO: 5 is V_(L1), SEQ ID NO: 6is V_(L2), SEQ ID NO: 7 is V_(L3), and SEQ ID NO: 8 is V_(L4).

TABLE 3 SEQ ID NO: Name Amino acid sequence 5 V_(L1)DVVMTQSPSFLSASVGDRVTITCKASQHVGTAVAWYQQRPGKAPKLLIYWASTRHTGVPSRFSGSGSGTEFTLT ISSLQPEDFATYFCQQYSSYPFTFGGGTKLEIK6 V_(L2) DIVMTQSPDSLAVSLGERATINCKASQHVGTAVAWYQQKPGQPPKLLIYWASTRHTGVPDRFSGSGSGTDFTLT ISSLQAEDVAVYYCQQYSSYPFTFGQGTKLEIK7 V_(L3) DIQMTQSPFSLSASVGDRVTITCKASQHVGTAVAWYQQKPGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLT ISSLQPEDFATYFCQQYSSYPFTFGGGTKLEIK8 V_(L4) DIVMTQSPDSLAVSLGERATINCKASQHVGTAVAWYQQKPEQPPKLLIYWASTRHTGVPDRFSGSGSGTDFTLT ISSLQAEDVAVYYCQQYSSYPFTFGGGTKVEIK

Each of the V_(H) domains can be synthesized in-frame with a constantdomain sequence, for example, a human IgG1, IgG2, IgG3, IgG4, IgE, IgA1,IgA2, IgM, or IgD sequence. A DNA sequence encoding the entire heavychain sequence can be codon optimized and verified.

Illustrative amino acid sequences of constant domain sequences areprovided in TABLE 4. In some embodiments, a V_(H) domain disclosedherein is synthesized in-frame with a human IgG1 constant domainsequence. A human IgG1 constant domain sequence can comprise SEQ ID NO:220. In some embodiments, a V_(H) domain disclosed herein is synthesizedin-frame with a human IgG2 constant domain sequence. A human IgG2constant domain sequence can comprise SEQ ID NO: 221. In someembodiments, a V_(H) domain disclosed herein is synthesized in-framewith a human IgG3 constant domain sequence. A human IgG3 constant domainsequence can comprise SEQ ID NO: 222. In some embodiments, a V_(H)domain disclosed herein is synthesized in-frame with a human IgG4constant domain sequence. The human IgG4 isotype constant domainsequence can be mutated to a proline rather than a serine at position228 to reduce Fab-arm exchange (stabilizing S228P mutation). An aminoacid sequence of the IgG4 constant domain with S228P mutation cancomprise SEQ ID NO: 9. In some embodiments, a V_(H) domain disclosedherein is synthesized in-frame with a human IgE constant domainsequence. A human IgE constant domain sequence can comprise SEQ ID NO:223. In some embodiments, a V_(H) domain disclosed herein is synthesizedin-frame with a human IgA1 constant domain sequence. A human IgA1constant domain sequence can comprise SEQ ID NO: 224. In someembodiments, a V_(H) domain disclosed herein is synthesized in-framewith a human IgA2 constant domain sequence. A human IgA2 constant domainsequence can comprise SEQ ID NO: 225. In some embodiments, a V_(H)domain disclosed herein is synthesized in-frame with a human IgMconstant domain sequence. A human IgM constant domain sequence cancomprise SEQ ID NO: 226. In some embodiments, a V_(H) domain disclosedherein is synthesized in-frame with a human IgD constant domainsequence. A human IgD constant domain sequence can comprise SEQ ID NO:227.

TABLE 4 SEQ ID NO: Name Amino acid sequence 220 IgG1ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS constantGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 221 IgG2ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNS constantGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDISVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGK222 IgG3 ASTKGPSVFPLAPCSRSTSGGTAALGCLVKDYFPEPVTVSWNS constantGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYTCNVNHKPSNTKVDKRVELKTPLGDTTHTCPRCPEPKSCDTPPPCPRCPEPKSCDTPPPCPRCPEPKSCDTPPPCPRCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFKWYVDGVEVHNAKTKPREEQYNSTFRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKLTVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPGK 9 IgG4ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNS constantGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK 223 IgEASTQSPSVFPLTRCCKNIPSNATSVTLGCLATGYFPEPVMVTW constantDTGSLNGTTMTLPATTLTLSGHYATISLLTVSGAWAKQMFTCRVAHTPSSTDWVDNKTFSVCSRDFTPPTVKILQSSCDGGGHFPPTIQLLCLVSGYTPGTINITWLEDGQVMDVDLSTASTTQEGELASTQSELTLSQKHWLSDRTYTCQVTYQGHTFEDSTKKCADSNPRGVSAYLSRPSPFDLFIRKSPTITCLVVDLAPSKGTVNLTWSRASGKPVNHSTRKEEKQRNGTLTVTSTLPVGTRDWIEGETYQCRVTHPHLPRALMRSTTKTSGPRAAPEVYAFATPEWPGSRDKRTLACLIQNFMPEDISVQWLHNEVQLPDARHSTTQPRKTKGSGFFVFSRLEVTRAEWEQKDEFICRAVHEAASPSQTVQRAVSVN PGK 224 IgA1ASPTSPKVFPLSLCSTQPDGNVVIACLVQGFFPQEPLSVTWSES constantGQGVTARNFPPSQDASGDLYTTSSQLTLPATQCLAGKSVTCHVKHYTNPSQDVTVPCPVPSTPPTPSPSTPPTPSPSCCHPRLSLHRPALEDLLLGSEANLTCTLTGLRDASGVTFTWTPSSGKSAVQGPPERDLCGCYSVSSVLPGCAEPWNHGKTFTCTAAYPESKTPLTATLSKSGNTFRPEVHLLPPPSEELALNELVTLTCLARGFSPKDVLVRWLQGSQELPREKYLTWASRQEPSQGTTTFAVTSILRVAAEDWKKGDTFSCMVGHEALPLAFTQKTIDRLAGKPTHVNV SVVMAEVDGTCY 225 IgGA2ASPTSPKVFPLSLDSTPQDGNVVVACLVQGFFPQEPLSVTWSE constantSGQNVTARNFPPSQDASGDLYTTSSQLTLPATQCPDGKSVTCHVKHYTNSSQDVTVPCRVPPPPPCCHPRLSLHRPALEDLLLGSEANLTCTLTGLRDASGATFTWTPSSGKSAVQGPPERDLCGCYSVSSVLPGCAQPWNHGETFTCTAAHPELKTPLTANITKSGNTFRPEVHLLPPPSEELALNELVTLTCLARGFSPKDVLVRWLQGSQELPREKYLTWASRQEPSQGTTTYAVTSILRVAAEDWKKGETFSCMVGHEALPLAFTQKTIDRMAGKPTHINVSVVMAEADGTC Y 226 IgMGSASAPTLFPLVSCENSPSDTSSVAVGCLAQDFLPDSITFSWKY constantKNNSDISSTRGFPSVLRGGKYAATSQVLLPSKDVMQGTDEHVVCKVQHPNGNKEKNVPLPVIAELPPKVSVFVPPRDGFFGNPRKSKLICQATGFSPRQIQVSWLREGKQVGSGVTTDQVQAEAKESGPTTYKVTSTLTIKESDWLGQSMFTCRVDHRGLTFQQNASSMCVPDQDTAIRVFAIPPSFASIFLTKSTKLTCLVTDLTTYDSVTISWTRQNGEAVKTHTNISESHPNATFSAVGEASICEDDWNSGERFTCTVTHTDLPSPLKQTISRPKGVALHRPDVYLLPPAREQLNLRESATITCLVTGFSPADVFVQWMQRGQPLSPEKYVTSAPMPEPQAPGRYFAHSILTVSEEEWNTGETYTCVVAHEALPNRVTE RTVDKSTGKPTLYNVSLVMSDTAGTCY227 IgD APTKAPDVFPIISGCRHPKDNSPVVLACLITGYHPTSVTVTWY constantMGTQSQPQRTFPEIQRRDSYYMTSSQLSTPLQQWRQGEYKCVVQHTASKSKKEIFRWPESPKAQASSVPTAQPQAEGSLAKATTAPATTRNTGRGGEEKKKEKEKEEQEERETKTPECPSHTQPLGVYLLTPAVQDLWLRDKATFTCFVVGSDLKDAHLTWEVAGKVPTGGVEEGLLERHSNGSQSQHSRLTLPRSLWNAGTSVTCTLNHPSLPPQRLMALREPAAQAPVKLSLNLLASSDPPEAASWLLCEVSGFSPPNILLMWLEDQREVNTSGFAPARPPPQPRSTTFWAWSVLRVPAPPSPQPATYTCVVSHEDSRTLLNASRSLEVSYVTD HGPMK

Each of the V_(L) domains can be synthesized in-frame with a human lightchain constant domain sequence, e.g. a kappa (IgK) or lambda (IgL)chain. The entire light chain sequence can then be codon optimized, andthe DNA sequence can be verified. TABLE 5 provides example light chainconstant domain sequences.

In some embodiments, a V_(L) domain disclosed herein is synthesizedin-frame with a human IgK constant domain sequence. A human IgK constantdomain sequence can comprise SEQ ID NO: 10. In some embodiments, a V_(L)domain disclosed herein is synthesized in-frame with a human IgLconstant domain sequence. A human IgL constant domain sequence cancomprise SEQ ID NO: 228.

TABLE 5 SEQ ID NO: Name Amino acid sequence 10 IgKTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY constantPREAKVQWKVDNALQSGNSQESVTEQDSKDST YSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 228 IgL GQPKANPTVTLFPPSSEELQANKATLVCLISD constantFYPGAVTVAWKADGSPVKAGVETTKPSKQSNN KYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS

Signal peptides can result in higher protein expression and/or secretionby a cell. The following signal peptides can be appended to all theconstructs disclosed herein.

Heavy chain signal peptide (SEQ ID NO: 11): MGWTLVFLFLLSVTAGVHSLight chain signal peptide (SEQ ID NO: 12): MVSSAQFLGLLLLCFQGTRC

Signal peptidases can cleave a signal peptide off a protein, forexample, during a secretion process, generating a mature protein thatdoes not comprise the signal peptide sequence. In some embodiments, asignal peptide is cleaved off a compound or antibody of the disclosure.In some embodiments, a mature compound or antibody of the disclosuredoes not comprise a signal peptide.

TABLE 6 and TABLE 7 list the full amino acid sequences of humanizedheavy and light chains that bind HPTP-β (VE-PTP), respectively. HC1,HC2, HC3 and HC4 are the human IgG4 isotype constant domain withstabilizing S228P mutation joined to V_(H1), V_(H2), V_(H3) and V_(H4),respectively, and with signal peptide appended. Amino acids 1-19 of SEQID NOs: 13, 14, 15, and 16 are the heavy chain signal peptide (SEQ IDNO: 11). Also shown are HC1, HC2, HC3, and HC4 without the signalpeptide appended (SEQ ID NOs: 246, 247, 248, and 249). In someembodiments, a mature compound or antibody of the disclosure does notcomprise the signal peptide(s).

LC1, LC2, LC3 and LC4 are the human IgK isotype constant domain joinedto V_(L1), V_(L2), V_(L3), and V_(L4), respectively, and with signalpeptide appended. Amino acids 1-20 of SEQ ID NOs: 17, 18, 19, and 20 arethe light chain signal peptide (SEQ ID NO: 12). Also shown are LC1, LC2,LC3, and LC4 without the signal peptide appended (SEQ ID NOs: 250, 251,252, and 253). In some embodiments, a mature compound or antibody of thedisclosure does not comprise the signal peptide(s).

TABLE 6 SEQ ID NO: Name Amino acid sequence 13 signalMGWTLVFLFLLSVTAGVHSEVQLVESGGGLVQPGGSLKLSCA peptide -ASGFTFNANAMNWVRQASGKGLEWVGRIRTKSNNYATYYAG HC1SVKDRFTISRDDSKNTAYLQMNSLKTEDTAAYYCVRDYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK 246 HC1EVQLVESGGGLVQPGGSLKLSCAASGFTFNANAMNWVRQASGKGLEWVGRIRTKSNNYATYYAGSVKDRFTISRDDSKNTAYLQMNSLKTEDTAAYYCVRDYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCS VMHEALHNHYTQKSLSLSLGK 14signal MGWTLVFLFLLSVTAGVHSEVQLVESGGGLVQPGGSLRLSCA peptide -ASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATYYAG HC2SVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRDYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK 247 HC2EVQLVESGGGLVQPGGSLRLSCAASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATYYAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRDYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVM HEALHNHYTQKSLSLSLGK 15signal MGWTLVFLFLLSVTAGVHSEVQLVESGGGLVQPGRSLRLSCT peptide -ASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATYYAG HC3SVKDRFTISRDDSKNIAYLQMNSLKTEDTAVYYCVRDYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK 248 HC3EVQLVESGGGLVQPGRSLRLSCTASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATYYAGSVKDRFTISRDDSKNIAYLQMNSLKTEDTAVYYCVRDYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVM HEALHNHYTQKSLSLSLGK 16signal MGWTLVFLFLLSVTAGVHSLVQLVESGGGLVKPGGSLRLSCA peptide -ASGFTFNANAMNWIRQAPGKGLEWVSRIRTKSNNYATYYAGS HC4VKDRFTISRDNAKNSLYLQMNSLRAEDTAVHYCVRDYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLEPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK 249 HC4LVQLVESGGGLVKPGGSLRLSCAASGFTENANAMNWIRQAPGKGLEWVSRIRTKSNNYATYYAGSVKDRFTISRDNAKNSLYLQMNSLRAEDTAVHYCVRDYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLEPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVM HEALHNHYTQKSLSLSLGK

TABLE 7 SEQ ID NO: Name Amino acid sequence 17 signalMVSSAQFLGLLLLCFQGTRCDVVMTQSPSFLSASVGDRVTIT peptide-CKASQHVGTAVAWYQQRPGKAPKLLIYWASTRHTGVPSRF LC1SGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGEC        250 LC1 DVVMTQSPSFLSASVGDRVTITCKASQHVGTAVAWYQQRPGKAPKLLIYWASTRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC 18 signalMVSSAQFLGLLLLCFQGTRCDIVMTQSPDSLAVSLGERATIN peptide-CKASQHVGTAVAWYQQKPGQPPKLLIYWASTRHTGVPDRF LC2SGSGSGTDFTLTISSLQAEDVAVYYCQQYSSYPFTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE KHKVYACEVTHQGLSSPVTKSFNRGEC251 LC2 DIVMTQSPDSLAVSLGERATINCKASQHVGTAVAWYQQKPGQPPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYSSYPFTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC 19 signalMVSSAQFLGLLLLCFQGTRCDIQMTQSPFSLSASVGDRVTIT peptide-CKASQHVGTAVAWYQQKPGKAPKLLIYWASTRHTGVPSRF LC3SGSGSGTDFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGEC 252LC3 DIQMTQSPFSLSASVGDRVTITCKASQHVGTAVAWYQQKPGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSF NRGEC 20 signalMVSSAQFLGLLLLCFQGTRCDIVMTQSPDSLAVSLGERATIN peptide-CKASQHVGTAVAWYQQKPEQPPKLLIYWASTRHTGVPDRF LC4SGSGSGTDFTLTISSLQAEDVAVYYCQQYSSYPFTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE KHKVYACEVTHQGLSSPVTKSFNRGEC253 LC4 DIVMTQSPDSLAVSLGERATINCKASQHVGTAVAWYQQKPEQPPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYSSYPFTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC

Any of the V_(H) regions disclosed herein can be combined with any ofthe V_(L) regions disclosed herein, with or without additional sequencesappended, to produce compounds that bind anti-HPTP-β (VE-PTP). Forexample, signal peptides and constant region sequences can be appendedto V_(H) and V_(L) regions as shown in TABLE 6 and TABLE 7 respectively,and the resulting heavy and light chains can be paired in anycombination to form antibodies. TABLE 8 shows 16 possible pairings ofHC1-4 and LC1-4 that bind HPTP-β (VE-PTP). Transfection and expressionof each of the anti-HPTP-β (VE-PTP) antibodies in TABLE 8 can bepursued.

TABLE 8 HC1:LC1 HC1:LC2 HC1:LC3 HC1:LC4 HC2:LC1 HC2:LC2 HC2:LC3 HC2:LC4HC3:LC1 HC3:LC2 HC3:LC3 HC3:LC4 HC4:LC1 HC4:LC2 HC4:LC3 HC4:LC4

Antibodies or antigen-binding compounds specific for HPTP-β (VE-PTP) canbe combined with antibodies or compounds that activate Tie2, inhibitVEGF, or inhibit VEGFR, to form multispecific compounds.

TABLE 9 provides sequences of collagen IV-derived biomimetic peptides.SEQ ID NO: 152 is AXT-107, an integrin-targeting collagen IV-derivedbiomimetic peptide that can inhibit VEGFRphosphorylation/activation/signaling and promote Tie2phosphorylation/activation/signaling. SEQ ID NO: 153 provides aconsensus sequence, wherein X is any standard amino acid ornon-genetically encoded amino acid. In some embodiments, X at position 7is M, A, or G; X at position 9 is F, A, Y, or G; X at position 0 is M,A, G, D-Alanine (dA), or norleucine (Nle); X at position 11 is F, A, Y,G, or 4-chlorophenylalanine (4-CiPhe): X at position 12 and position 18are independently selected from 2-Aminobutyric acid (Abu), G, S, A, V,T, I, L, or Allylglycine (AllylGly).

TABLE 9 SEQ ID NO: Amino acid sequence 152 LRRFSTAPFAFIDINDVINF 153LRRFSTXPXXXXNINNVXNF

TABLE 10 provides sequences related to vasculotide, a synthetic Ang1mimetic peptide that can act as a Tie2 agonist and activate Tie2signaling. SEQ ID NO: 229 provides the sequence of a synthetic 7-merthat binds the Tie2 receptor. SEQ ID NO: 230 provides an 8-mer with acysteine residue added at the N-terminus, allowing covalent tethering toa polyethylene glycol backbone to generate a tetrameric polyethyleneoxide clustered version of the peptide.

TABLE 10 SEQ ID NO: Amino acid sequence 229 HHHRHSF 230 CHHHRHSF

Antibodies or antigen-binding compounds specific for HPTP-β (VE-PTP) canbe combined with antibodies or compounds specific to VEGF or VEGFR toform multispecific compounds that bind HPTP-β (VE-PTP) and VEGF orVEGFR. TABLE 11, TABLE 12, TABLE 13, TABLE 14, TABLE 15, and TABLE 17provide example sequences of compounds specific for VEGF. TABLE 16provides sequences of an antibody that binds VEGFR.

TABLE 11 provides sequences related to aflibercept, a recombinantprotein comprising VEGF-binding portions of human VEGF receptors 1 and 2fused to the Fc portion of human IgG1. SEQ ID NO: 21 is the full aminoacid sequence of aflibercept. SEQ ID NO: 22 is a shortened sequencecontaining the VEGF-binding portions of human VEGF receptors 1 and 2without the Fc portion of IgG.

TABLE 11 SEQ ID NO: Name Amino acid sequence 21 AFL₁SDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEKDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHN HYTQKSLSLSPG 22 AFL₂SDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMT KKNSTFVRVHEK

TABLE 12 provides the sequence of brolucizumab (SEQ ID NO: 23), ahumanized single-chain antibody fragment (scFv) inhibitor of VEGF thatbinds to the receptor binding site of VEGF and thereby interferes withthe interaction of VEGF with VEGFR1 and VEGFR2.

TABLE 12 SEQ ID NO: Name Amino acid sequence 23 BRO_(scFv1)EIVMTQSPSTLSASVGDRVIITCQASEIIHSWLAWYQQKPGKAPKLLIYLASTLASGVPSRFSGSGSGAEFTLTISSLQPDDFATYYCQNVYLASTNGANFGQGTKLTVLGGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCTASGFSLTDYYYMTWVRQAPGKGLEWVGFIDPDDDPYYATWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCA GGDHNSGWGLDIWGQGTLVTVSS

TABLE 13 provides sequences related to ranibizumab, a humanizedmonoclonal antibody fragment (Fab) that binds to and inhibits activityof VEGF. SEQ ID NO: 24 is the heavy chain sequence of ranibizumab. SEQID NO: 25 is the light chain sequence of ranibizumab. SEQ ID NO: 26 is ashortened sequence of the ranibizumab heavy chain that can be used incloning a single-chain antibody fragment (scFv). SEQ ID NO: 27 is ashortened sequence of the ranibizumab light chain that can be used incloning a single-chain antibody fragment (scFv). SEQ ID NO: 28 is asingle-chain antibody fragment (scFv) comprising SEQ ID NO: 26, SEQ IDNO: 27, and SEQ ID NO: 33 (linker peptide, underlined).

TABLE 13 SEQ ID NO: Name Amino acid sequence 24 RAN_(H1)EVQLVESGGGLVQPGGSLRLSCAASGYDFTHYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPYYYGTSHWYFDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSCDKTHL 25 RAN_(L1)DIQLTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSTVPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC26 RAN_(H2) EVQLVESGGGLVQPGGSLRLSCAASGYDFTHYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPYYYGTSHW YFDVWGQGTLVTVSS 27 RAN_(L2)DIQLTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFSGSGSGTDFT LTISSLQPEDFATYYCQQYSTVPWTFGQGTKVEIK28 RAN_(scFv1) EVQLVESGGGLVQPGGSLRLSCAASGYDFTHYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPYYYGTSHWYFDVWGQGTLVTVSSGGGGSGGGGSGGGGSDIQLTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSL QPEDFATYYCQQYSTVPWTFGQGTKVEIK

TABLE 14 provides sequences related to bevacizumab, a humanizedmonoclonal antibody that that binds to and inhibits activity of VEGF.SEQ ID NO: 29 is the heavy chain sequence of bevacizumab. SEQ ID NO: 30is the light chain sequence of bevacizumab.

TABLE 14 SEQ ID NO: Name Amino acid sequence 29 BEV_(H1)EVQLVESGGGLVQPGGSLRLSCAASGYTFTNYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPHYYGSSHWYFDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGK 30 BEV_(L1)DIQMTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSTVPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSF NRGEC

TABLE 15 provides sequences related to conbercept, a recombinant proteincomprising extracellular domains from VEGF receptors 1 and 2 fused tothe Fc portion of human IgG1. SEQ ID NO: 154 is the full amino acidsequence of conbercept. SEQ ID NO: 155 is a shortened sequencecontaining sequences from VEGF receptors 1 and 2 without the Fc portionof IgG.

TABLE 15 SEQ ID NO: Name Amino acid sequence 154 CON₁GRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEKPFVAFGSGMESLVEATVGERVRIPAKYLGYPPPEIKWYKNGIPLESNHTIKAGHVLTIMEVSERDTGNYTVILTNPISKEKQSHVVSLVVYVPPGPGDKTHTCPLCPAPELLGGPSVFLEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKATPPVLDSDGSFFLYSKLTVDKSRWQQ GNVFSCSVMHEALHNHYTQKSLSLSPGK155 CON₂ GRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEKPFVAFGSGMESLVEATVGERVRIPAKYLGYPPPEIKWYKNGIPLESNHTIKAGHVLTIMEVSERDTGNYTVILTNPISKEKQSHVVSLVVY VPPGPG

TABLE 16 provides sequences related to ramucirumab, a humanizedmonoclonal antibody that binds to an extracellular domain of VEGFR2 andinhibits VEGFR2 signaling. SEQ ID NO: 156 is the heavy chain sequence oframucirumab. SEQ ID NO: 157 is the light chain sequence of ramucirumab.

TABLE 16 SEQ ID NO: Name Amino acid sequence 156 RAM_(H1)EVQLVQSGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSSISSSSSYIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARVTDAFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL SLSPGK 157 RAM_(L1)DIQMTQSPSSVSASIGDRVTITCRASQGIDNWLGWYQQKPGKAPKLLIYDASNLDTGVPSRFSGSGSGTYFTLTISSLQAEDFAVYFCQQAKAFPPTFGGGTKVDIKGTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG LSSPVTKSFNRGEC

TABLE 17 provides DARPin and DAR-Pin-derived amino acid sequences thatbind VEGF and inhibit VEGFR signaling. SEQ ID NOS: 158-168 comprisedesigned ankyrin repeats with binding specificity for VEGF. SEQ ID NO:169 comprises designed ankyrin repeats with a binding specificity forVEGF and designed ankyrin repeats with a binding specificity for serumalbumin. SEQ ID NO: 170 comprises designed ankyrin repeats with abinding specificity for VEGF, designed ankyrin repeats with a bindingspecificity for hepatocyte growth factor, and designed ankyrin repeatswith a binding specificity for serum albumin. SEQ ID NOS: 171-177comprise designed ankyrin repeats with a binding specificity for VEGF.SEQ ID NO: 173 provides the sequence of abicipar, which comprisesdesigned ankyrin repeats with a binding specificity for VEGF. SEQ IDNOS: 178-190 provide individual designed ankyrin repeat sequence motifswith binding specificity for VEGF, wherein X represents any amino acid.SEQ ID NOS: 191-217 comprise designed ankyrin repeats with bindingspecificity for VEGF.

TABLE 17 SEQ ID NO: Amino acid sequence 158DLGKKLLEAARAGQDDEVRILMANGADVNAFDWMGWTPLHLAAHEGHLEIVEVLLKNGADVNATDVSGYTPLHLAAADGHLEIVEVLLKHGADVNTKDNTGWTPLHLSADLGHLEIVEVLLKNGADVNAQDKFGKTAF DISIDNGNEDLAEILQKAA 159DLDKKLLEAARAGQDDEVRILLKAGADVNAKDYLGWTPLHLAAHEGHLEIVEVLLKAGADVNAKDVSGYTPLHLAAADGHLEIVEVLLKAGADVNAKDNTGWTPLHLSADLGHLEIVEVLLKAGADVNAQDKFGKTAFDI SIDNGNEDLAEILQKAA 160DLGKKLLEAARAGQDDEVRILMANGADVNTADSTGWTPLHLAAPWGHPEIVEVLLKNGADVNAHDYQGWTPLHLAATLGHLEIVEVLLKHGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA 161DLGKKLLEAARAGQDDEVRILMANGADVNTADSTGWTPLHLAVPWGHLEIVEVLLKYGADVNAKDFQGWTPLHLAAAIGHQEIVEVLLKNGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA 162DLDKKLLEAARAGQDDEVRILMANGADVNAKDSTGYTPLHLAAPWGHLEIVEVLLKAGADVNAKDYQGWTPLHLAAAVGHLEIVEVLLKAGADVNAQDKSGKTPADLAADAGHEDIAEVLQKAA 163DLGKKLLEAARAGQDDEVRILMANGADVNARDSTGWTPLHLAAPWGHPEIVEVLLKNGADVNAADFQGWTPLHLAAAVGHLEIVEVLLKHGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA 164DLDKKLLEAARAGQDDEVRILLKAGADVNAKDSTGWTPLHLAAPWGHPEIVEVLLKAGADVNAKDFQGWTPLHLAAAAGHLEIVEVLLKAGADVNAQDKSGKTPADLAADAGHEDIAEVLQKAA 165DLGKKLLEAARAGQDDEVRILLKAGADVNAKDSTGWTPLHLAAPWGHPEIVEVLLKAGADVNAKDFQGWTPLHLAAAAGHLEIVEVLLKAGADVNAQDKSGKTPADLAADAGHEDIAEVLQKAA 166DLDKKLLEAARAGQDDEVRILLKAGADVNAKDSTGWTPLHLAAPWGHPEIVEVLLKAGADVNAKDFQGWTPLHLAAAVGHLEIVEVLLKAGADVNAQDKSGKTPADLAADAGHEDIAEVLQKAA 167DLDKKLLEAARAGQDDEVRILLKAGADVNAKDSTGWTPLHLAAPWGHPEIVEVLLKAGADVNAKDYQGWTPLHLAAAVGHLEIVEVLLKAGADVNAQDKSGKTPADLAADAGHEDIAEVLQKAA 168DLDKKLLEAARAGQDDEVRILMANGADVNAKDSTGWTPLHLAAPWGHLEIVEVLLKAGADVNAKDFQGWTPLHLAAAVGHLEIVEVLLKAGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA 169DLGKKLLEAARAGQDDEVRELLKAGADVNAKDYFSHTPLHLAARNGHLKIVEVLLKAGADVNAKDFAGKTPLHLAANEGHLEIVEVLLKAGADVNAQDIFGKTPADIAADAGHEDIAEVLQKAAGSPTPTPTTPTPTPTTPTPTPTGSDLDKKLLEAARAGQDDEVRILLKAGADVNAKDSTGWTPLHLAAPWGHPEIVEVLLKAGADVNAKDFQGWTPLHLAAAAGHLEIVEVLLKAGADVNAQDKSGKTPADLAADAGHEDIAEVLQKAA 170GSDLGKKLLEAARAGQDDEVRELLKAGADVNAKDYFSHTPLHLAARNGHLKIVEVLLKAGADVNAKDFAGKTPLHLAANEGHLEIVEVLLKAGADVNAQDIFGKTPADIAADAGHEDIAEVLQKAAGSPTPTPTTPTPTPTTPTPTPTGSDLGKKLLEAARAGQDDEVRILLKAGADVNAKDRYGDTPLHLAADIGHLEIVEVLLKAGADVNAEDYFGNTPLHLAASYGHLEIVEVLLKAGADVNAKDDYGNTPLHLAANTGHLEIVEVLLKAGADVNAQDKSGKTPADLAADAGHEDIAEVLQKAAGSPTPTPTTPTPTPTTPTPTPTGSDLDKKLLEAARAGQDDEVRILLKAGADVNAKDSTGWTPLHLAAPWGHPEIVEVLLKAGADVNAKDFQGWTPLHLAAAAGHLEIVEVLLKAGADVNAQDKSGKTPADLAADAGHEDIAEVLQKAAGSPTPTPTTPTPTPTTPTPTPTGSDLGKKLLEAARAGQDDEVRELLKAGADVNAKDYFSHTPLHLAARNGHLKIVEVLLKAGADVNAKDFAGKTPLHLAANEGHLEIVEVLLKAGADVNAQDIFGKTPADIAADAGHEDIAEVLQKAA 171GSDLGKKLLEAARAGQDDEVRILMANGADVNAFDWMGWTPLHLAAHEGHLEIVEVLLKNGADVNATDVSGYTPLHLAAADGHLEIVEVLLKYGADVNTKDNTGWTPLHLSADLGRLEIVEVLLKYGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAASGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPGSEPATSGSETPGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPSEGSAPGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPGTSTEPSEGSAPGTSTEPSEGSAPGTSESATPESGPGTSESATPESGPGSPAGSPTSTEEGTSESATPESGPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPGTSESATPESGPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGSPAGSPTSTEEGSPAGSPTSTEEGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGTSESATPESGPGSEPATSGSETPGTSESATPESGPGSEPATSGSETPGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTEEGTSESATPESGPGSEPATSGSETPGTSESATPESGPGSPAGSPTSTEEGSPAGSPTSTEEGTSTEPSEGSAPGTSESATPESGPGTSESATPESGPGTSESATPESGPGSEPATSGSETPGSEPATSGSETPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPSEGSAPGSEPATSGSETPGTSES ATPESGPGTSTEPSEGSAPG172 GSDLGKKLLEAARAGQDDEVRILMANGADVNAFDWMGWTPLHLAAHEGHLEIVEVLLKNGADVNATDVSGYTPLHLAAADGHLEIVEVLLKHGADVNTKDNTGWTPLHLSADLGHLEIVEVLLKNGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAAGGGSGGGSC 173GSDLDKKLLEAARAGQDDEVRILMANGADVNARDSTGWTPLHLAAPWGHPEIVEVLLKNGADVNAADFQGWTPLHLAAAVGHLEIVEVLLKYGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAAGGGSGGGSC 174GSDLGKKLLEAARAGQDDEVRILMANGADVNTADSTGWTPLHLAVPWGHLEIVEVLLKYGADVNAKDFQGWTPLHLAAAIGHQEIVEVLLKNGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAAGSGSASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASPAAPAPASPAAPAPSAPAASP AAPAPAS 175GSDLGKKLLEAARVGQDDEVRILMADGADVNASDFKGDTPLHLAASQGHLEIVEVLLKYGADVNAYDMLGWTPLHLAADLGHLEIVEVLLKYGADVNAQDRFGKTAFDISIDNGNEDLAEILQKAAGSPSTADGC 176GSDLGKKLLEAVRAGQDDEVRILMTNGADVNAKDQFGFTPLQLAAYNGHLEIVEVLLKYGADVNAFDIFGWTPLHLAADLGHLEIVEVLLKNGADVNAQDKFGRTAFDISIDNGNEDLAEILQKAASGSC 177GSDLGKKLLEAARAGQDDEVRILMANGADVNAVDYIGWTPLHLAAAYGHLEIVEVLLKYSADVNAEDFAGYTPLHLAASNGHLEIVEVLLKYGADVNTKDNTGWTPLHLSADLGHLEIVEVLLKYGADVNTQDKFGKTAFDISIDNGNEDLAEILQKAAGSPSTADGC 178 XDXXGXTPLHLAAXXGHLEIVEVLLKXGADVNA 179XDXXGWTPLHLAAXXGHLEIVEVLLKXGADVNA 180 XDXXGXTPLHLAAXXGHLEIVEVLLKXGADVNX181 XDXXGWTPLHLXADLGXLEIVEVLLKXGADVNX 182XDXXGXTPLHLAAXXGHXEIVEVLLKXGADVNA 183 XDXXGXTPLHLAAXXGHLEIVEVLLKXGADVNA184 XDXXGWTPLHLAAXXGHLEIVEVLLKXGADVNA 185XDXXGXTPLHLAAXXGHLEIVEVLLKXGADVNX 186 XDXXGXTPLHLXAXXGHLEIVEVLLKXGADVNA187 XDFKXDTPLHLAAXXGHXEIVEVLLKXGADVNA 188XDXLXXTPLHLAXXXGHLEIVEVLLKXGADVNA 189 XDXXGXTPLXLAAXXGHLEIVEVLLKXGADVNA190 XDXXGWTXLHLAADLGXLEIVEVLLKXGADVNA 191GSDLGKKLLEAARAGQDDEVRILMANGADVNAFDWMGWTPLHLAAHEGHLEIVEVLLKNGADVNATDVSGYTPLHLAAADGHLEIVEVLLKYGADVNTKDNTGWTPLHLSADLGHLEIVEVLLKYGADVNAQDKFGKT AFDISIDNGNEDLAEILQKAA 192GSDLGKKLLEAARVGQDDEVRILMANGADVNAFDWMGWTPLHLAAHEGHLEIVEVLLKNGADVNATDVSGYTPLHLAAADGHLEIVEVLLKYGADVNTKDNTGWTPLHLSADLGHLEIVEVLLKYGADVNAQDKFGKT AFDISIDNGNEDLAEILQKAA 193GSDLGKKLLEAARAGQDDEVRILMANGADVNAFDWMGWTPLHLAAHEGHLEIVEVLLKNGADVNATDVSGYTPLHLAAADGHLEIVEVLLKYGADVNTKDNTGWTPLHLSADLGRLEIVEVLLKYGADVNAQDKFGKT AFDISIDNGNEDLAEILQKAA 194GSDLGKKLLEAARAGQDDEVRILMANGADVNAFDWMGWTPLHLAAHEGHLEIVEVLLKNGTDVNATDVSGYTPLHLAAADGHLEIVEVLLKYGADVNTKDNTGWTPLHLSADLGHLEIVEVLLKHGADVNAQDKFGKT AFDISIDNGNEDLAEILQKAA 195GSDLGKKLLEAARAGQDDEVRILMANGADVNAFDWMGWTPLHLAAHEGHLEIVEVLLKNGADVNATDVSGYTPLHLAAADGHLEIVEVLLKHGADVNTKDNTGWTPLHLSADLGHLEIVEVLLKNGADVNAQDKFGKT AFDISIDNGNEDLAEILQKAA 196GSDLGKKLLEAARAGQDDEVRILMANGADVNAFDWMGWTPLHLAAHEGHLEIVEVLLKNGADVNATDVSGYTPLHLAAADGHLEIVEVLLKHGADVNTKDNTGWTPLHLSADLGHLEIVEVLLKNGADINAQDKFGKTA FDISIDNGNEDLAEILQKAA 197GSDLGKKLLEAARAGQDDEVRILMANGADVNAFDWMGWTPLHLAAHEGHLEIVEVLLKNGADVNATDVSGYTPLHLAAADGHLEIVEVLLKHGADVNTTDNTGWTPLHLSADLGHLEIVEVLLKYGADVNAQDKFGKT AFDISIDNGNEDLAEILQKAA 198GSDLGKKLLEAARAGQDDEVRILMANGADVNAFDYMGWTPLHLAAHNGHMEIVEVLLKYGADVNASDYSGYTPLHLAAADGHLEIVEVLLKYGADVNTKDNTGWTPLHLSADLGHLEIVEVLLKYGADVNAQDKFGKT AFDISIDNGNEDLAEILQKAA 199GSDLGKKLLEAARAGQDDEVRILMANGADVNAVDYIGWTPLHLAAAYGHLEIVEVLLKYSADVNAEDFAGYTPLHLAASNGHLEIVEVLLKYGADVNTKDNTGWTPLHLSADLGHLEIVEVLLKYGADVNTQDKFGKTAFD ISIDNGNEDLAEILQKAA 200GSDLGKKLLEAARTGQDDEVRILMANGADVNATDYMGWTPLHLAAKVGHLEIVEVLLKYGADVNAEDYNGYTPLHLAAAMGHLEIAEVLLKYGADVNTKDNTGWTPLHLSADLGHLEIVEVLLKNGADVNAQDKFGKT AFDISIDNGNEDLAEILQKAA 201GSDLGKKLLEAARAGQDDEVRILMANGADVNARDSTGWTPLHLAAPWGHPEIVEVLLKNGADVNAADFQGWTPLHLAAAVGHLEIVEVLLKYGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA 202GSDLGKKLLEAARAGQDDEVRILMANGADVNARDSTGWTPLHLAAPWGHPEIVEVLLKNGADVNAADFQGWTPLHLAAAVGHLEIVEVLLKHGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA 203GSDLGKKLLEAARAGQDDEVRILMANGADVNTADSTGWTPLHLAAPWGHPEIVEVLLKNGADVNAHDYQGWTPLHLAATLGHLEIVEVLLKYGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA 204GSDLGKKLLEAARAGQDDEVRILMANGADVNTADSTGWTPLHLVAPWGHPEIVEVLLKHGADVNTHDYQGWTPLHLAATLGHLEIVEVLLRYGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA 205GSDLGKKLLEAARAGQDDEVRILMANGADVNTADSTGWTPMHLAAPWGHPEIVEVLLKHGADVNAQDFQGWTPLHLAAAIGHLEIVEVLLKYGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA 206GSDLGKKLLEAARAGQDDEVRILMANGADVNTADSTGWTPLHLAVPWGHLEIVEVLLKYGADVNAKDFQGWTPLHLAAAIGHQEIVEVLLKNGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA 207GSDLGKKLLEAARVGQDDEVRILMADGADVNASDFKGDTPLHLAASQGHLEIVEVLLKYGADVNAYDMLGWTPLHLAADLGHLEIVEVLLKYGADVNAQDRFGKTAFDISIDNGNEDLAEILQKAA 208GSDLGKKLLEAARVGQDDEVRILMANGADVNASDFKGDTPLHLAASQGHLEIVEVLLKNSADVNAFDLLGWTPLHLAADLGHLEIVEVLLKYGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA 209GSDLGKKLLEAARVGQDDEVRILMANGADVNALDFKGDTPLHLAAASGHLEIVEVLLKNGADVNAHDMLSWTPLHLAGDLGHLEIVEVLLKYGADVNAQDRFGKTAFDISIDNGNEDLAEILQKAA 210GSDLGKKLLEAVRAGQDDEVRILMTNGADVNAKDQFGFTPLQLAAYNGHLEIVEVLLKYGADVNAFDIFGWTPLHLAADLGHLEIVEVLLKNGADVNAQDKFGRTAFDISIDNGNEDLAEILQKAA 211GSDLGKKLLEAVRAGQDDEVRILMANGADVNASDNQGTTPLHLAASHGHLEIVEVLLKYGADVNDAHDDLGWTPLHLSADLGHLEIVEVLLKYGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA 212GSDLGKKLLEATRAGQDDEVRILMANGADVNASDNQGTTPLHLAASHGHLEIVEVLLKYGADVNDAHDDLGWTPLHLAADLGHLEIVEVLLKYGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA 213GSDLGKKLLEAARVGQDDEVRILMADGADVNASDFKGDTPLHLAASQGHLEIVEVLLKYGADVNAYDMLGWTPLHLAADLGHLEIVEVLLKYGADVNAQDRFGKTAFDISIDNGNEDLAEILQKAA 214GSDLGKKLLEAARVGQDDEVRILMANDADVNASDFKGDTPLHLAASQGHLEIVEVLLKYGADVNAYDMLGWTPLHLAADLGHLEIVEVLLKHGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA 215GSDLGKKLLEAARAGQDDEVRILMANGADVNTLDFKSDTPLHLAAASGHLEIVEVLLKNGADVNAHDMLSWTPLHLAGDLGHLEIVEVLLKHGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA 216GSDLGKKLLEAARAGQDDEVRILMANGADVNAKDIYGRTPLHLAALHGHPEIVEVLLKYGADVNANDYWGTTSLHLVAIWGHLEIVEVLLKYGADVNAVDDIGQTPLHLAAAWGHLEIVEVLLKHGADVNAQDKFGKTA FDISIDNGNEDLAEILQKAA 217GSDLGKKLLEAARAGQDDEVRILMANGADVNANDYDGMTPLHLAAMEGHLEIVEVLLKYGADVNANDHYGFTPLHLAWTGRLEIVEVLLKNGADVNAADVFGRTPLHLAATSGHLEIVEVLLKYGADVNAQDKFGKTAF DISIDNGNEDLAEILQKAA

Antibodies or antigen-binding compounds specific for VEGF can becombined with antibodies or antigen-binding compounds specific to HPTP-β(VE-PTP) to form multispecific compounds, such as bispecific compoundsthat bind VEGF and HPTP-β (VE-PTP). Any compounds in this disclosurespecific for VEGF can be combined with any compounds in this disclosurespecific to HPTP-β (VE-PTP). Any of the compounds in this disclosurespecific for VEGF or HPTP-β (VE-PTP) can be modified as necessary forthe generation of a multispecific compound. Non-limiting examples ofmodifications necessary for the generation of a multispecific compoundinclude the addition of amino acid residues, the removal of amino acidresidues, the replacement of amino acid residues, and the use oflinkers. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or more residues can beadded to an N-terminus and/or a C-terminus of a sequence disclosedherein, and the resulting sequence can be used in the generation of amulti-specific construct. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, ormore residues can be removed from an N-terminus and/or a C-terminus of asequence disclosed herein, and the remaining sequence can be used in thegeneration of a multi-specific construct. For example, N- and/orC-terminal residues can be removed from SEQ ID NO: 173 (e.g., togenerate SEQ ID NO: 244), and the truncated sequence can be used in amulti-specific construct. In some embodiments, the sequences in any ofSEQ ID NOS: 13-20 or 246-253 can be modified. For example, one or moreC-terminal residues can be removed (e.g., the C-terminal lysine can beremoved from any of SEQ ID NOS: 13-16 or 246-249, and the remainingresidues (residues 1-467) can be used in a multi-specific construct).

A compound described herein can include a linker between differentdomains of the compound. A linker can be a chemical bond, for example, acovalent bond or a non-covalent bond. A linker as described herein caninclude a flexible or rigid linker.

A linker of the disclosure can include a chemical linker. For example,two amino acid sequences of the disclosure can be connected together bya chemical linker. Each chemical linker of the disclosure can bealkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene,heterocycloalkylene, arylene, or heteroarylene, any of which isoptionally substituted. In some embodiments, a chemical linker of thedisclosure can be an ester, ether, amide, thioether, orpolyethyleneglycol (PEG). In some embodiments, a linker can reverse theorder of the amino acids sequence in a compound, for example, so thatthe amino acid sequences linked by the linked are head-to-head, ratherthan head-to-tail. Non-limiting examples of such linkers includediesters of dicarboxylic acids, such as oxalyl diester, malonyl diester,succinyl diester, glutaryl diester, adipyl diester, pimetyl diester,fumaryl diester, maleyl diester, phthalyl diester, isophthalyl diester,and terephthalyl diester. Non-limiting examples of such linkers includediamides of dicarboxylic acids, such as oxalyl diamide, malonyl diamide,succinyl diamide, glutaryl diamide, adipyl diamide, pimetyl diamide,fumaryl diamide, maleyl diamide, phthalyl diamide, isophthalyl diamide,and terephthalyl diamide. Non-limiting examples of such linkers includediamides of diamino linkers, such as ethylene diamine,1,2-di(methylamino)ethane, 1,3-diaminopropane,1,3-di(methylamino)propane, 1,4-di(methylamino)butane,1,5-di(methylamino)pentane, 1,6-di(methylamino)hexane, and pipyrizine.

Non-limiting examples of optional substituents include hydroxyl groups,sulfhydryl groups, halogens, amino groups, nitro groups, nitroso groups,cyano groups, azido groups, sulfoxide groups, sulfone groups,sulfonamide groups, carboxyl groups, carboxaldehyde groups, iminegroups, alkyl groups, halo-alkyl groups, alkenyl groups, halo-alkenylgroups, alkynyl groups, halo-alkynyl groups, alkoxy groups, aryl groups,aryloxy groups, aralkyl groups, arylalkoxy groups, heterocyclyl groups,acyl groups, acyloxy groups, carbamate groups, amide groups, ureidogroups, epoxy groups, and ester groups.

A linker can be a peptide. A linker can comprise a linker sequence, forexample, a linker peptide sequence. A linker sequence can be, forexample, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36,37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 51, 53, 54,55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, or 70 aminoacid residues in length.

A flexible linker can have a sequence containing stretches of glycineand serine residues. The small size of the glycine and serine residuesprovides flexibility, and allows for mobility of the connectedfunctional domains. The incorporation of serine or threonine canmaintain the stability of the linker in aqueous solutions by forminghydrogen bonds with the water molecules, thereby reducing unfavorableinteractions between the linker and protein moieties.

Flexible linkers can also contain additional amino acids such asthreonine and alanine to maintain flexibility, as well as polar aminoacids such as lysine and glutamine to improve solubility.

A flexible linker can comprise repeats of SEQ ID NO: 42 (GGGS), forexample, SEQ ID NOS: 42-55. A flexible linker can comprise repeats ofSEQ ID NO: 31 (GGGGS), for example, SEQ ID NOS: 31-41. Several othertypes of flexible linkers, including SEQ ID NO: 59 (KESGSVSSEQLAQFRSLD)and SEQ ID NO: 60 (EGKSSGSGSESKST), can also be used. The SEQ ID NO: 61(GSAGSAAGSGEF) linker can also be used, in which large hydrophobicresidues are minimized to maintain good solubility in aqueous solutions.The length of the flexible linkers can be adjusted to allow for properfolding or to achieve optimal biological activity of the fused proteins.

A rigid linker can have, for example, an alpha helix-structure. Analpha-helical rigid linker can act as a spacer between protein domains.A rigid linker can comprise repeats of SEQ ID NO: 62 (EAAAK), forexample, SEQ ID NOS: 62-66. A rigid linker can comprise repeats of SEQID NO: 67 (EAAAR), for example, SEQ ID NOS: 67-72. A rigid linker canhave a proline-rich sequence, (XP)n, with X designating alanine, lysine,glutamine, or any amino acid. The presence of proline in non-helicallinkers can increase stiffness, and allow for effective separation ofprotein domains.

A linker can comprise any of the sequences disclosed in TABLE 18, whichcan be used to link any portion of a compound disclosed herein to anyportion of another compound disclosed herein:

TABLE 18 SEQ  ID NO: Sequence 31 GGGGS 32 GGGGSGGGGS 33 GGGGSGGGGSGGGGS34 GGGGSGGGGSGGGGSGGGGS 35 GGGGSGGGGSGGGGSGGGGSGGGGS 36GGGGSGGGGSGGGGSGGGGSGGGGSGGGGS 37 GGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGS 38GGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGS 39GGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGS 40GGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGS 41GGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGS 42 GGGS 43GGGSGGGS 44 GGGSGGGSGGGS 45 GGGSGGGSGGGSGGGS 46 GGGSGGGSGGGSGGGSGGGS 47GGGSGGGSGGGSGGGSGGGSGGGS 48 GGGSGGGSGGGSGGGSGGGSGGGSGGGS 49GGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGS 50 GGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGS51 GGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGS 52GGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGS 53GGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGS 54GGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGS 55GGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGSGGGS 56 GG 57 GGGGGG58 GGGGGGGG 59 KESGSVSSEQLAQFRSLD 60 EGKSSGSGSESKST 61 GSAGSAAGSGEF 62EAAAK 63 EAAAKEAAAK 64 EAAAKEAAAKEAAAKEAAAK 65 EAAAKEAAAKEAAAKEAAAKEAAAK66 EAAAKEAAAKEAAAKEAAAKEAAAKEAAAK 67 EAAAR 68 EAAAREAAAR 69EAAAREAAAREAAAR 70 EAAAREAAAREAAAREAAAR 71 EAAAREAAAREAAAREAAAREAAAR 72EAAAREAAAREAAAREAAAREAAAREAAAR 73AEAAAKEAAAKEAAAKEAAAKALEAEAAAKEAAAKEAAAKEAAAKA 74 PAPAP 75 AEAAAKEAAAKA

The VEGF-binding and VEGFR-binding compounds described in TABLE 11,TABLE 12, TABLE 13, TABLE 14, TABLE 15, TABLE 16, and TABLE 17 can becombined with the HPTP-β (VE-PTP)-binding compounds described in TABLE2, TABLE 3, TABLE 6, TABLE 7, and TABLE 8.

Any of the 16 antibodies described in TABLE 8 can be combined withaflibercept or aflibercept-related sequences to generate a bispecificantibody. Non-limiting schematic examples are presented in FIG. 2 , FIG.3 , and FIG. 4 . For example, SEQ ID NO: 22 can be appended onto SEQ IDNO: 14 or SEQ ID NO: 247 of antibody HC2:LC1 to generate a tetravalentbispecific antibody wherein the heavy chain of HC2:LC1 is appended withSEQ ID NO: 22 (FIG. 2 ). SEQ ID NO: 22 can be appended onto SEQ ID NO:17 or SEQ ID NO: 250 of antibody HC2:LC1 to generate a tetravalentbispecific antibody, in which the light chain of HC2:LC1 is appendedwith SEQ ID NO: 22 (FIG. 3 ). SEQ ID NO: 22 can be appended onto SEQ IDNO: 14 or SEQ ID NO: 247, and SEQ ID NO: 22 can be appended onto SEQ IDNO: 17 or SEQ ID NO: 250 of antibody HC2:LC1 to generate a hexavalentbispecific antibody wherein the heavy and light chains of HC2:LC1 areappended with SEQ ID NO: 22 (FIG. 4 ).

Any of the 16 antibodies described in TABLE 8 can be combined withbrolucizumab or brolucizumab-related sequences to generate a bispecificantibody. Non-limiting schematic examples are presented in FIG. 2 , FIG.3 , and FIG. 4 . For example, SEQ ID NO: 23 can be appended onto SEQ IDNO: 14 or SEQ ID NO: 247 of antibody HC2:LC1 to generate a tetravalentbispecific antibody wherein the heavy chain of HC2:LC1 is appended withSEQ ID NO: 23 (FIG. 2 ). SEQ ID NO: 23 can be appended onto SEQ ID NO:17 or SEQ ID NO: 250 of antibody HC2:LC1 to generate a tetravalentbispecific antibody wherein the light chain of HC2:LC1 is appended withSEQ ID NO: 23 (FIG. 3 ). SEQ ID NO: 23 can be appended onto SEQ ID NO:14 or SEQ ID NO: 247, and SEQ ID NO: 23 can be appended onto SEQ ID NO:17 or SEQ ID NO: 250 of antibody HC2:LC1 to generate a hexavalentbispecific antibody wherein the heavy and light chains of HC2:LC1 areappended with SEQ ID NO: 23 (FIG. 4 ).

Any of the 16 antibodies described in TABLE 8 can be combined withranibizumab or ranibizumab-related sequences to generate a bispecificantibody. Non-limiting schematic examples are presented in FIG. 2 , FIG.3 , and FIG. 4 . For example, SEQ ID NO: 28 can be appended onto SEQ IDNO: 14 or SEQ ID NO: 247 of antibody HC2:LC1 to generate a tetravalentbispecific antibody wherein the heavy chain of HC2:LC1 is appended withSEQ ID NO: 28 (FIG. 2 ). SEQ ID NO: 28 can be appended onto SEQ ID NO:17 or SEQ ID NO: 250 of antibody HC2:LC1 to generate a tetravalentbispecific antibody wherein the light chain of HC2:LC1 is appended withSEQ ID NO: 28 (FIG. 3 ). SEQ ID NO: 28 can be appended onto SEQ ID NO:14 or SEQ ID NO: 247, and SEQ ID NO: 28 can be appended onto SEQ ID NO:17 or SEQ ID NO: 250 of antibody HC2:LC1 to generate a hexavalentbispecific antibody wherein the heavy and light chains of HC2:LC1 areappended with SEQ ID NO: 28 (FIG. 4 ).

Any of the 16 antibodies described in TABLE 8 can be combined withbevacizumab or bevacizumab-related sequences to generate a bispecificantibody. Non-limiting schematic examples are presented in FIG. 2 , FIG.3 , and FIG. 4 . For example, an antigen-binding scFv of bevacizumabcould be generated as demonstrated for ranibizumab in TABLE 13. Thebevacizumab-derived antigen-binding scFv can be appended onto SEQ ID NO:14 or SEQ ID NO: 247 of antibody HC2:LC1 to generate a tetravalentbispecific antibody wherein the heavy chain of HC2:LC1 is appended withthe bevacizumab-derived antigen-binding scFv (FIG. 2 ). Thebevacizumab-derived antigen-binding scFv can be appended onto SEQ ID NO:17 or SEQ ID NO: 250 of antibody HC2:LC1 to generate a tetravalentbispecific antibody, in which the light chain of HC2:LC1 is appendedwith the bevacizumab-derived antigen-binding scFv (FIG. 3 ). Thebevacizumab-derived antigen-binding scFv can be appended onto SEQ ID NO:14 or SEQ ID NO: 247, and the bevacizumab-derived antigen-binding scFvcan be appended onto SEQ ID NO: 17 or SEQ ID NO: 250 of antibody HC2:LC1to generate a hexavalent bispecific antibody, in which the heavy andlight chains of HC2:LC1 are appended with the bevacizumab-derivedantigen-binding scFv (FIG. 4 ).

Any of the 16 antibodies described in TABLE 8 can be combined withabicipar or abicipar-related sequences to generate a bispecificantibody. Non-limiting schematic examples are presented in FIG. 2 , FIG.3 , and FIG. 4 . For example, SEQ ID NO: 173 or SEQ ID NO: 244 can beappended onto SEQ ID NO: 14, SEQ ID NO: 247, residues 1-467 SEQ ID NO:14, or residues 1-467 SEQ ID NO: 247 of antibody HC2:LC1 to generate atetravalent bispecific antibody wherein the heavy chain of HC2:LC1 isappended with SEQ ID NO: 173 or SEQ ID NO: 244 (FIG. 2 ). SEQ ID NO: 173or SEQ ID NO: 244 can be appended onto SEQ ID NO: 17 or SEQ ID NO: 250of antibody HC2:LC1 to generate a tetravalent bispecific antibody, inwhich the light chain of HC2:LC1 is appended with SEQ ID NO: 173 or SEQID NO: 244 (FIG. 3 ). SEQ ID NO: 173 or SEQ ID NO: 244 can be appendedonto SEQ ID NO: 14, SEQ ID NO: 247, residues 1-467 SEQ ID NO: 14, orresidues 1-467 SEQ ID NO: 247, and SEQ ID NO: 173 or SEQ ID NO: 244 canbe appended onto SEQ ID NO: 17 or SEQ ID NO: 250 of antibody HC2:LC1 togenerate a hexavalent bispecific antibody wherein the heavy and lightchains of HC2:LC1 are appended with SEQ ID NO: 173 or SEQ ID NO: 244(FIG. 4 ).

Any of the 16 antibodies described in TABLE 8 can be combined withconbercept or conbercept-related sequences to generate a bispecificantibody. Non-limiting schematic examples are presented in FIG. 2 , FIG.3 , and FIG. 4 . For example, SEQ ID NO: 155 can be appended onto SEQ IDNO: 14 or SEQ ID NO: 247 of antibody HC2:LC1 to generate a tetravalentbispecific antibody wherein the heavy chain of HC2:LC1 is appended withSEQ ID NO: 155 (FIG. 2 ). SEQ ID NO: 155 can be appended onto SEQ ID NO:17 or SEQ ID NO: 250 of antibody HC2:LC1 to generate a tetravalentbispecific antibody, in which the light chain of HC2:LC1 is appendedwith SEQ ID NO: 155 (FIG. 3 ). SEQ ID NO: 155 can be appended onto SEQID NO: 14 or SEQ ID NO: 247, and SEQ ID NO: 155 can be appended onto SEQID NO: 17 or SEQ ID NO: 250 of antibody HC2:LC1 to generate a hexavalentbispecific antibody wherein the heavy and light chains of HC2:LC1 areappended with SEQ ID NO: 155 (FIG. 4 ).

Any of the 16 antibodies described in TABLE 8 can be combined withramucirumab or ramucirumab-related sequences to generate a bispecificantibody. Non-limiting schematic examples are presented in FIG. 2 , FIG.3 , and FIG. 4 . For example, an antigen-binding scFv of ramucirumabcould be generated as demonstrated for ranibizumab in TABLE 13. Theramucirumab-derived antigen-binding scFv can be appended onto SEQ ID NO:14 or SEQ ID NO: 247 of antibody HC2:LC1 to generate a tetravalentbispecific antibody wherein the heavy chain of HC2:LC1 is appended withthe ramucirumab-derived antigen-binding scFv (FIG. 2 ). Theramucirumab-derived antigen-binding scFv can be appended onto SEQ ID NO:17 or SEQ ID NO: 250 of antibody HC2:LC1 to generate a tetravalentbispecific antibody, in which the light chain of HC2:LC1 is appendedwith the ramucirumab-derived antigen-binding scFv (FIG. 3 ). Theramucirumab-derived antigen-binding scFv can be appended onto SEQ ID NO:14 or SEQ ID NO: 247, and the ramucirumab-derived antigen-binding scFvcan be appended onto SEQ ID NO: 17 or SEQ ID NO: 250 of antibody HC2:LC1to generate a hexavalent bispecific antibody, in which the heavy andlight chains of HC2:LC1 are appended with the ramucirumab-derivedantigen-binding scFv (FIG. 4 ).

Any of the 16 antibodies described in TABLE 8 can be combined withDARPins, DARPin repeats, or sequences therefrom, to generate amultispecific antibody. Non-limiting schematic examples are presented inFIG. 2 , FIG. 3 , and FIG. 4 . For example, an amino acid sequencecomprising any of SEQ ID NOS: 158-217 can be appended onto SEQ ID NO: 14or SEQ ID NO: 247 of antibody HC2:LC1 to generate a tetravalentbispecific antibody wherein the heavy chain of HC2:LC1 is appended withan amino acid sequence comprising any of SEQ ID NOS: 158-217 (FIG. 2 ).An amino acid sequence comprising any of SEQ ID NOS: 158-217 can beappended onto SEQ ID NO: 17 or SEQ ID NO: 250 of antibody HC2:LC1 togenerate a tetravalent bispecific antibody, in which the light chain ofHC2:LC1 is appended with an amino acid sequence comprising any of SEQ IDNOS: 158-217 (FIG. 3 ). An amino acid sequence comprising any of SEQ IDNOS: 158-217 can be appended onto SEQ ID NO: 14 or SEQ ID NO: 247 andonto SEQ ID NO: 17 or SEQ ID NO: 250 of antibody HC2:LC1 to generate ahexavalent bispecific antibody wherein the heavy and light chains ofHC2:LC1 are appended with an amino acid sequence comprising any of SEQID NOS: 158-217 (FIG. 4 ).

The Tie2 activating compounds described in TABLE 9 and TABLE 10 can becombined with the HPTP-β (VE-PTP)-binding compounds described in TABLE2, TABLE 3, TABLE 6, TABLE 7, and TABLE 8.

Any of the 16 antibodies described in TABLE 8 can be combined withcollagen IV-derived biomimetic peptide sequences to generate amultispecific antibody. Non-limiting schematic examples are presented inFIG. 2 , FIG. 3 , and FIG. 4 . For example, an amino acid sequencecomprising SEQ ID NO: 152 or SEQ ID NO: 153 can be appended onto SEQ IDNO: 14 or SEQ ID NO: 247 of antibody HC2:LC1 to generate a tetravalentbispecific antibody wherein the heavy chain of HC2:LC1 is appended withan amino acid sequence comprising SEQ ID NO: 152 or SEQ ID NO: 153 (FIG.2 ). An amino acid sequence comprising SEQ ID NO: 152 or SEQ ID NO: 153can be appended onto SEQ ID NO: 17 or SEQ ID NO: 250 of antibody HC2:LC1to generate a tetravalent bispecific antibody, in which the light chainof HC2:LC1 is appended with an amino acid sequence comprising SEQ ID NO:152 or SEQ ID NO: 153 (FIG. 3 ). An amino acid sequence comprising SEQID NO: 152 or SEQ ID NO: 153 can be appended onto SEQ ID NO: 14 or SEQID NO: 247 and onto SEQ ID NO: 17 or SEQ ID NO: 250 of antibody HC2:LC1to generate a hexavalent bispecific antibody wherein the heavy and lightchains of HC2:LC1 are appended with an amino acid sequence comprisingSEQ ID NO: 152 or SEQ ID NO: 153 (FIG. 4 ).

Any of the 16 antibodies described in TABLE 8 can be combined with Ang2mimetics, or sequences therefrom, to generate a multispecific antibody.Non-limiting schematic examples are presented in FIG. 2 , FIG. 3 , andFIG. 4 . For example, an amino acid sequence comprising any of SEQ IDNOS: 229-230 can be appended onto SEQ ID NO: 14 or SEQ ID NO: 247 ofantibody HC2:LC1 to generate a tetravalent bispecific antibody whereinthe heavy chain of HC2:LC1 is appended with an amino acid sequencecomprising any of SEQ ID NOS: 229-230 (FIG. 2 ). An amino acid sequencecomprising any of SEQ ID NOS: 229-230 can be appended onto SEQ ID NO: 17or SEQ ID NO: 250 of antibody HC2:LC1 to generate a tetravalentbispecific antibody, in which the light chain of HC2:LC1 is appendedwith an amino acid sequence comprising any of SEQ ID NOS: 229-230 (FIG.3 ). An amino acid sequence comprising any of SEQ ID NOS: 229-230 can beappended onto SEQ ID NO: 14 or SEQ ID NO: 247 and onto SEQ ID NO: 17 orSEQ ID NO: 250 of antibody HC2:LC1 to generate a hexavalent bispecificantibody wherein the heavy and light chains of HC2:LC1 are appended withan amino acid sequence comprising any of SEQ ID NOS: 229-230 (FIG. 4 ).

The Tie2 activating compounds described in TABLE 9 and TABLE 10 can becombined with the VEGF-binding and VEGFR-binding compounds described inTABLE 11, TABLE 12, TABLE 13, TABLE 14, TABLE 15, TABLE 16, and TABLE 17to generate multi-specific compounds.

Any of the compounds in this disclosure, e.g., the multi-specific fusionconstructs described above, can be modified as necessary to promotedesirable protein folding or biological activity.

CDRs

Sequences in this disclosure can comprise complementarity determiningregions (CDRs). CDRs can be identified by the Kabat method, the Chothiamethod, the IMGT method, or the Paratome method. A CDR of a sequenceherein can be, for example, between 0 and 91 residues in length, between0 and 25 residues in length, between 5 and 14 residues in length, about0 residues in length, about 1 residue in length, about 2 residues inlength, about 3 residues in length, about 4 residues in length, about 5residues in length, about 6 residues in length, about 7 residues inlength, about 8 residues in length, about 9 residues in length, about 10residues in length, about 11 residues in length, about 12 residues inlength, about 13 residues in length, about 14 residues in length, about15 residues in length, about 16 residues in length, about 17 residues inlength, about 18 residues in length, about 19 residues in length, about20 residues in length, about 21 residues in length, about 22 residues inlength, about 23 residues in length, about 24 residues in length, orabout 25 residues in length.

A compound of this disclosure with a binding specificity for or anability to modulate HPTP-β (VE-PTP) can comprise, for example, any ofthe CDRs in TABLE 19, TABLE 20, TABLE 21, TABLE 22, TABLE 23, or TABLE24.

TABLE 19 provides non-limiting examples of HCDR1 sequences specific forHPTP-β (VE-PTP).

TABLE 19 SEQ ID NO: Amino acid sequence 76 ANAMN 77 GFTFNAN 78 GFTFNANA79 FTFNANAMN 80 GFTFNANAMN

TABLE 20 provides non-limiting examples of HCDR2 sequences specific forHPTP-β (VE-PTP).

TABLE 20 SEQ ID NO: Amino acid sequence 81 RIRTKSNNYATYYAGSVKD 82RTKSNNYA 83 IRTKSNNYAT 84 WVGRIRTKSNNYATYY 85 WVGRIRTKSNNYATYYAGSVKD 86WVSRIRTKSNNYATYY 87 WVSRIRTKSNNYATYYAGSVKD

TABLE 21 provides non-limiting examples of HCDR3 sequences specific forHPTP-β (VE-PTP).

TABLE 21 SEQ ID NO: Amino acid sequence 88 DYYGSSAWITY 89 VRDYYGSSAWITY90 RDYYGSSAWITY

TABLE 22 provides non-limiting examples of LCDR1 sequences specific forHPTP-β (VE-PTP).

TABLE 22 SEQ ID NO: Amino acid sequence 91 KASQHVGTAVA 92 QHVGTA 93QHVGTAVA

TABLE 23 provides non-limiting examples of LCDR2 sequences specific forHPTP-β (VE-PTP).

TABLE 23 SEQ ID NO: Amino acid sequence 94 WASTRHT 95 WAS 96 LLIYWASTRHT

TABLE 24 provides non-limiting examples of LCDR3 sequences specific forHPTP-β (VE-PTP).

TABLE 24 SEQ ID NO: Amino acid sequence 97 QQYSSYPFT 98 QQYSSYPF

Compounds of this disclosure with a binding specificity for or anability to modulate VEGF can comprise any of the CDRs in TABLE 25, TABLE26, TABLE 27, TABLE 28, TABLE 29, or TABLE 30.

TABLE 25 provides non-limiting examples of HCDR1 sequences specific forVEGF.

TABLE 25 SEQ ID NO: Amino acid sequence 99 DYYYMT 100 GFSLTDYY 101GFSLTDYYY 102 FSLTDYYYMT 103 GFSLTDYYYMT 104 HYGMN 105 GYDFTHY 106GYDFTHYG 107 YDFTHYGMN 108 GYDFTHYGMN 109 NYGMN 110 GYTFTNY 111 GYTFTNYG112 YTFTNYGMN 113 GYTFTNYGMN

TABLE 26 provides non-limiting examples of HCDR2 sequences specific forVEGF.

TABLE 26 SEQ ID NO: Amino acid sequence 114 FIDPDDDPYYATWAKG 115 DPDDD116 IDPDDDP 117 WVGFIDPDDDPYYATWA 118 WVGFIDPDDDPYYATWAKG 119WINTYTGEPTYAADFKR 120 NTYTGE 121 INTYTGEP 122 WVGWINTYTGEPTY 123WVGWINTYTGEPTYAADFKR

TABLE 27 provides non-limiting examples of HCDR3 sequences specific forVEGF.

TABLE 27 SEQ ID NO: Amino acid sequence 124 GDHNSGWGLDI 125AGGDHNSGWGLDI 126 YPYYYGTSHWYFDV 127 AKYPYYYGTSHWYFDV 128KYPYYYGTSHWYFDV 129 YPHYYGSSHWYFDV 130 AKYPHYYGSSHWYFDV 131KYPHYYGSSHWYFDV

TABLE 28 provides non-limiting examples of LCDR1 sequences specific forVEGF.

TABLE 28 SEQ ID NO: Amino acid sequence 132 QASEIIHSWLA 133 EIIHSW 134EIIHSWLA 135 SASQDISNYLN 136 QDISNY 137 QDISNYLN

TABLE 29 provides non-limiting examples of LCDR2 sequences specific forVEGF.

TABLE 29 SEQ ID NO: Amino acid sequence 138 LASTLAS 139 LAS 140LLIYLASTLAS 141 FTSSLHS 142 FTS 143 VLIYFTSSLHS

TABLE 30 provides non-limiting examples of LCDR3 sequences specific forVEGF.

TABLE 30 SEQ ID NO: Amino acid sequence 144 QNVYLASTNGAN 145 QQYSTVPWT146 QQYSTVPW

TABLE 31 provides aflibercept-derived sequences corresponding to the D2domain of human VEGF receptor 1 (SEQ ID NO: 147) and the D3 domain ofhuman VEGF receptor 2 (SEQ ID NO: 148).

TABLE 31 SEQ  ID NO: Amino acid sequence 147SDTGRPFVEMYSEIPEIIHMTEGRELVIPCR VTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYL THRQTNTII 148DVVLSPSHGIELSVGEKLVLNCTARTELNVG IDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKK NSTFVRVHEK

TABLE 32 provides abicipar-derived sequences. SEQ ID NOS: 233-236correspond to ankyrin repeats within abicipar. SEQ ID NOS: 237-242provide consensus sequences for VEGF binding ankyrin repeats. In SEQ IDNO: 237, X1 is K, T, or Y; X2 is N or M; X3 is T or F; X4 is S or A; X5is H or R; X6 is A, Y, H, or N; and X7 is A or T. In SEQ ID NO: 238, X1is K, M, N, R, or V; X2 is Y, H, M, or V; X3 is F, L, M, or V; X4 is R,H, V, A, K, or N; X5 is F, D, H, T, Y, M, or K; and X6 is A, H, N, or Y.In SEQ ID NO: 239, X1 is L, S, or T; X2 is G, S, or C; X3 is S or A; X4is Q, S, M, or N; X5 is L, M, or Q; and X6 is A, H, N, Y, or D. In SEQID NO: 240, X1 is K, S, N, T, or V; X2 is K, N, W, A, H, M, Q, or S; X3is F, Q, L, H, or V; X4 is F or T; X5 is Q or H; X6 is Y or S; X7 is N,H, Y, or M; and X8 is A, H, N, or Y. In SEQ ID NO: 241, X1 is A, N, R,V, Y, E, H, I, K, L, Q, S, or T; X2 is S, A, N, R, D, F, L, P, T, or Y;X3 is T, V, S, A, L, or F; X4 is W, F, or H; X5 is P, I, A, L, S, T, V,or Y; X6 is W, F, I, L, T, or V; X7 is L or P; and X8 is A, H, N, or Y.In SEQ ID NO: 242, X1 is H, Q, A, K, R, D, I, L, M, N, V, or Y; X2 is Y,F, or H; X3 is Q, F, or T; X4 is W, M, G, H, N, or T; X5 is T, A, M, L,or V; X6 is I, L, V, D, or T; and X7 is A, H, N, or Y. SEQ ID NO: 244provides a truncated sequence derived from abicipar that comprisesdesigned ankyrin repeats with a binding specificity for VEGF.

TABLE 32 SEQ  ID NO: Amino acid sequence 233DLDKKLLEAARAGQDDEVRILMANGADVNARDS 234 TGWTPLHLAAPWGHPEIVEVLLKNGADVNAADF235 QGWTPLHLAAAVGHLEIVEVLLKYGADVNAQDK 236 FGKTAFDISIDNGNEDLAEILQ 237X1DX2X3GWTPLHLX4ADLGX5LEIVEVLLKX6GADVNX7 238X1DX2X3GWTPLHLAAX4X5GHLEIVEVLLKX6GADVNA 239X1DFKX2DTPLHLAAX3X4GHX5EIVEVLLKX6GADVNA 240X1DX2X3GX4TPLX5LAAX6X7GHLEIVEVLLKX8GADVNA 241X1DX2X3GX4TPLHLAAX5X6GHX7EIVEVLLKX8GADVNA 242X1DX2X3GX4TPLHLAAX5X6GHLEIVEVLLKX7GADVNADLDKKLLEAARAGQDDEVRILMANGADVNARDST GWTPLHLAAPWGHPEIVE 244VLLKNGADVNAADFQGWTPLHLAAAVGHLEIVEVLLKYGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAAHomology

A sequence of a compound herein can have at least about 70% homology, atleast about 71% homology, at least about 72% homology, at least about73% homology, at least about 74% homology, at least about 75% homology,at least about 76% homology, at least about 77% homology, at least about78% homology, at least about 79% homology, at least about 80% homology,at least about 81% homology, at least about 82% homology, at least about83% homology, at least about 84% homology, at least about 85% homology,at least about 86% homology, at least about 87% homology, at least about88% homology, at least about 89% homology, at least about 90% homology,at least about 91% homology, at least about 92% homology, at least about93% homology, at least about 94% homology, at least about 95% homology,at least about 96% homology, at least about 97% homology, at least about98% homology, at least about 99% homology, at least about 99.1%homology, at least about 99.2% homology, at least about 99.3% homology,at least about 99.4% homology, at least about 99.5% homology, at leastabout 99.6% homology, at least about 99.7% homology, at least about99.8% homology, at least about 99.9% homology, at least about 99.91%homology, at least about 99.92% homology, at least about 99.93%homology, at least about 99.94% homology, at least about 99.95%homology, at least about 99.96% homology, at least about 99.97%homology, at least about 99.98% homology, or at least about 99.99%homology to an amino acid sequence provided herein.

Various methods and software programs can be used to determine thehomology between two or sequences, such as NCBI BLAST, Clustal W, MAFFT,Clustal Omega, AlignMe, Praline, or another suitable method oralgorithm.

Pharmaceutical Compositions

A pharmaceutical composition of the disclosure can be a combination ofany pharmaceutical compounds described herein with other chemicalcomponents, such as carriers, stabilizers, diluents, dispersing agents,suspending agents, thickening agents, and/or excipients. Thepharmaceutical composition facilitates administration of the compound toan organism.

Pharmaceutical formulations for administration can include aqueoussolutions of the active compounds in water-soluble form. Suspensions ofthe active compounds can be prepared as oily injection suspensions.Suitable lipophilic solvents or vehicles include fatty oils such assesame oil, or synthetic fatty acid esters, such as ethyl oleate ortriglycerides, or liposomes. Aqueous injection suspensions can containsubstances which increase the viscosity of the suspension, such assodium carboxymethyl cellulose, sorbitol, or dextran. The suspension canalso contain suitable stabilizers or agents which increase thesolubility of the compounds to allow for the preparation of highlyconcentrated solutions. The active ingredient can be in powder form forconstitution with a suitable vehicle, for example, sterile pyrogen-freewater, before use.

In practicing the methods of treatment or use provided herein,therapeutically-effective amounts of the compounds described herein areadministered in pharmaceutical compositions to a subject having adisease or condition to be treated. In some embodiments, the subject isa mammal such as a human. A therapeutically-effective amount can varywidely depending on the severity of the disease, the age and relativehealth of the subject, the potency of the compounds used, and otherfactors.

Pharmaceutical compositions can be formulated using one or morephysiologically-acceptable carriers comprising excipients andauxiliaries, which facilitate processing of the active compounds intopreparations that can be used pharmaceutically. Formulation can bemodified depending upon the route of administration chosen.Pharmaceutical compositions comprising compounds described herein can bemanufactured, for example, by mixing, dissolving, emulsifying,encapsulating, entrapping, or compression processes.

The pharmaceutical compositions can include at least onepharmaceutically-acceptable carrier, diluent, or excipient and compoundsdescribed herein as free-base or pharmaceutically-acceptable salt form.Pharmaceutical compositions can contain solubilizers, stabilizers,tonicity enhancing agents, buffers, and preservatives.

Methods for the preparation of compositions comprising the compoundsdescribed herein include formulating the compounds with one or moreinert, pharmaceutically-acceptable excipients or carriers to form asolid, semi-solid, or liquid composition. Solid compositions include,for example, powders, tablets, dispersible granules, capsules, andcachets. Liquid compositions include, for example, solutions in which acompound is dissolved, emulsions comprising a compound, or a solutioncontaining liposomes, micelles, or nanoparticles comprising a compoundas disclosed herein. Semi-solid compositions include, for example, gels,suspensions and creams. The compositions can be in liquid solutions orsuspensions, solid forms suitable for solution or suspension in a liquidprior to use, or as emulsions. These compositions can also contain minoramounts of nontoxic, auxiliary substances, such as wetting oremulsifying agents, pH buffering agents, and otherpharmaceutically-acceptable additives.

Non-limiting examples of dosage forms suitable for use in the disclosureinclude liquid, powder, gel, nanosuspension, nanoparticle, microgel,aqueous or oily suspensions, emulsion, and any combination thereof.

Non-limiting examples of pharmaceutically-acceptable excipients suitablefor use in the disclosure include binding agents, disintegrating agents,anti-adherents, anti-static agents, surfactants, anti-oxidants, coatingagents, coloring agents, plasticizers, preservatives, suspending agents,emulsifying agents, anti-microbial agents, spheronization agents, andany combination thereof.

A composition of the disclosure can be, for example, an immediaterelease form or a controlled release formulation. An immediate releaseformulation can be formulated to allow the compounds to act rapidly.Non-limiting examples of immediate release formulations include readilydissolvable formulations. A controlled release formulation can be apharmaceutical formulation that has been adapted such that release ratesand release profiles of the active agent can be matched to physiologicaland chronotherapeutic requirements, or has been formulated to effectrelease of an active agent at a programmed rate. Non-limiting examplesof controlled release formulations include granules, delayed releasegranules, hydrogels (e.g., of synthetic or natural origin), othergelling agents (e.g., gel-forming dietary fibers), matrix-basedformulations (e.g., formulations comprising a polymeric material havingat least one active ingredient dispersed through), granules within amatrix, polymeric mixtures, and granular masses.

In some, a controlled release formulation is a delayed release form. Adelayed release form can be formulated to delay a compound's action foran extended period of time. A delayed release form can be formulated todelay the release of an effective dose of one or more compounds, forexample, for about 4, about 8, about 12, about 16, or about 24 hours.

A controlled release formulation can be a sustained release form. Asustained release form can be formulated to sustain, for example, thecompound's action over an extended period of time. A sustained releaseform can be formulated to provide an effective dose of any compounddescribed herein (e.g., provide a physiologically-effective bloodprofile) over about 4, about 8, about 12, about 16, or about 24 hours.

The disclosed compositions can optionally comprisepharmaceutically-acceptable preservatives.

Non-limiting examples of pharmaceutically-acceptable excipients can befound, for example, in Remington: The Science and Practice of Pharmacy,Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, JohnE., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton,Pennsylvania 1975; Liberman, H. A. and Lachman, L., Eds., PharmaceuticalDosage Forms, Marcel Decker, New York, N.Y., 1980; and PharmaceuticalDosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams& Wilkins 1999), each of which is incorporated by reference in itsentirety.

A compound described herein can be conveniently formulated intopharmaceutical compositions composed of one or morepharmaceutically-acceptable carriers. See e.g., Remington'sPharmaceutical Sciences, latest edition, by E.W. Martin Mack Pub. Co.,Easton, PA, incorporated by reference in its entirety, which disclosestypical carriers and conventional methods of preparing pharmaceuticalcompositions. Such carriers can be carriers for administration ofcompositions to humans and non-humans, including solutions such assterile water, saline, and buffered solutions at physiological pH.Pharmaceutical compositions can also include one or more additionalactive ingredients such as antimicrobial agents, anti-inflammatoryagents, and anesthetics.

Non-limiting examples of pharmaceutically-acceptable carriers includesaline, Ringer's solution, and dextrose solution. In some embodiments,the pH of the solution can be from about 5 to about 8, and can be fromabout 7 to about 7.5. Further carriers include sustained releasepreparations such as semipermeable matrices of solid hydrophobicpolymers containing the compound. The matrices can be in the form ofshaped articles, for example, films, liposomes, microparticles, ormicrocapsules.

Compositions suitable for topical administration can be used. In someembodiments, compositions of the disclosure can comprise a liquidcomprising an active agent in solution, in suspension, or both. Liquidcompositions can include gels. A liquid composition can be, for example,aqueous. A composition is an in situ gellable aqueous composition. Initeration, the composition is an in situ gellable aqueous solution. Sucha composition can comprise a gelling agent in a concentration effectiveto promote gelling upon contact with the eye or lacrimal fluid in theexterior of the eye. Aqueous compositions can have ophthalmically-compatible pH and osmolality. The composition can comprise anophthalmic depot formulation comprising an active agent forsubconjunctival administration. Microparticles comprising an activeagent can be embedded in a biocompatible, pharmaceutically-acceptablepolymer or a lipid encapsulating agent. The depot formulations can beadapted to release all or substantially all the active material over anextended period of time. The polymer or lipid matrix, if present, can beadapted to degrade sufficiently to be transported from the site ofadministration after release of all or substantially all the activeagent. The depot formulation can be a liquid formulation, comprising apharmaceutical acceptable polymer and a dissolved or dispersed activeagent. Upon injection, the polymer forms a depot at the injections site,for example, by gelifying or precipitating. The composition can comprisea solid article that can be inserted in a suitable location in the eye,such as between the eye and eyelid or in the conjuctival sac, where thearticle releases the active agent. Solid articles suitable forimplantation in the eye in such fashion can comprise polymers and can bebioerodible or non-bioerodible.

Pharmaceutical formulations can include additional carriers, as well asthickeners, diluents, buffers, preservatives, and surface active agentsin addition to the agents disclosed herein.

The pH of the disclosed composition can range from about 3 to about 12.The pH of the composition can be, for example, from about 3 to about 4,from about 4 to about 5, from about 5 to about 6, from about 6 to about7, from about 7 to about 8, from about 8 to about 9, from about 9 toabout 10, from about 10 to about 11, or from about 11 to about 12 pHunits. The pH of the composition can be, for example, about 3, about 4,about 5, about 6, about 7, about 8, about 9, about 10, about 11, orabout 12 pH units. The pH of the composition can be, for example, atleast 3, at least 4, at least 5, at least 6, at least 7, at least 8, atleast 9, at least 10, at least 11 or at least 12 pH units. The pH of thecomposition can be, for example, at most 3, at most 4, at most 5, atmost 6, at most 7, at most 8, at most 9, at most 10, at most 11, or atmost 12 pH units. A pharmaceutical formulation disclosed herein can havea pH of from about 5.5 to about 6.5. For example, a formulation of thepresent disclosure can have a pH of about 5.5, about 5.6, about 5.7,about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, about6.4, or about 6.5. In some embodiments, the pH is 6.2±0.3, 6.2±0.2,6.2±0.1, about 6.2, or 6.2.

If the pH is outside the range desired by the formulator, the pH can beadjusted by using sufficient pharmaceutically-acceptable acids andbases.

Depending on the intended mode of administration, the pharmaceuticalcompositions can be in the form of solid, semi-solid or liquid dosageforms, such as, for example, tablets, suppositories, pills, capsules,powders, liquids, suspensions, lotions, creams, or gels, for example, inunit dosage form suitable for single administration of a precise dosage.

For solid compositions, nontoxic solid carriers include, for example,pharmaceutical grades of mannitol, lactose, starch, magnesium stearate,sodium saccharin, talc, cellulose, glucose, sucrose, and magnesiumcarbonate.

Non-limiting examples of pharmaceutically active agents suitable forcombination with compositions of the disclosure include anti-infectives,i.e., aminoglycosides, antiviral agents, antimicrobials,anti-cholinergics/anti-spasmotics, antidiabetic agents, antihypertensiveagents, anti-neoplastics, cardiovascular agents, central nervous systemagents, coagulation modifiers, hormones, immunologic agents,immunosuppressive agents, and ophthalmic preparations.

In some embodiments, the pharmaceutical composition provided hereincomprises a therapeutically effective amount of a compound in admixturewith a pharmaceutically-acceptable carrier and/or excipient, forexample, saline, phosphate buffered saline, phosphate and amino acids,polymers, polyols, sugar, buffers, preservatives, and other proteins.Illustrative agents include octylphenoxy polyethoxy ethanol compounds,polyethylene glycol monostearate compounds, polyoxyethylene sorbitanfatty acid esters, sucrose, fructose, dextrose, maltose, glucose,mannitol, dextran, sorbitol, inositol, galactitol, xylitol, lactose,trehalose, bovine or human serum albumin, citrate, acetate, Ringer's andHank's solutions, cysteine, arginine, carnitine, alanine, glycine,lysine, valine, leucine, polyvinylpyrrolidone, polyethylene, and glycol.

In some embodiments, a pharmaceutical formulation disclosed herein cancomprise: (i) a compound or antibody disclosed herein; (ii) a buffer;(iii) a non-ionic detergent; (iv) a tonicity agent; and (v) astabilizer. In some embodiments, the pharmaceutical formulationdisclosed herein is a stable liquid pharmaceutical formulation.

In some embodiments, an ophthalmic formulation disclosed herein cancomprise: (i) a compound or antibody disclosed herein; (ii) a buffer;(iii) a non-ionic detergent; (iv) a tonicity agent; and (v) astabilizer. In some embodiments, the ophthalmic formulation disclosedherein is a stable liquid pharmaceutical formulation or a stable liquidophthalmic formulation.

In some embodiments, a pharmaceutical formulation or ophthalmicformulation disclosed herein is a liquid formulation that can compriseabout 5 mg/mL to about 150 mg/mL of antibody or compound, about 7.5mg/mL to about 140 mg/mL of antibody or compound, about 10 mg/mL toabout 130 mg/mL of antibody or compound, about 10 mg/mL to about 100mg/mL of antibody or compound, about 20 mg/mL to about 80 mg/mL ofantibody or compound, or about 30 mg/mL to about 70 mg/mL of antibody orcompound. For example, a formulation of the present disclosure cancomprise about 5 mg/mL, about 10 mg/mL, about 15 mg/mL, about 20 mg/mL,about 25 mg/mL, about 30 mg/mL, about 35 mg/mL, about 40 mg/mL, about 50mg/mL, about 60 mg/mL, about 70 mg/mL, about 80 mg/mL, about 90 mg/mL,about 100 mg/mL, about 120 mg/mL, about 140 mg/mL, or about 150 mg/mL ofa compound, antibody, or antigen-binding fragment thereof describedherein.

In some embodiments, a pharmaceutical formulation or ophthalmicformulation disclosed herein can comprise a buffer. In some embodiments,the buffer serves to maintain a stable pH and to help stabilize acompound or antibody disclosed herein. In some embodiments, the bufferor buffer system comprises at least one buffer that has a bufferingrange that overlaps fully or in part the range of pH 5.5-7.4. In someembodiments, the buffer has a pKa of about 6.2±0.5. In some embodiments,the buffer comprises a sodium phosphate buffer. In some embodiments, thesodium phosphate is present at a concentration of about 5 mM to about 15mM, about 6 mM to about 14 mM, about 7 mM to about 13 mM, about 8 mM toabout 12 mM, about 9 mM to about 11 mM, or about 10 mM. In certainembodiments, the buffer system comprises sodium phosphate at 10 mM, at apH of 6.20.3 or 6.10.3.

In some embodiments, a pharmaceutical formulation or ophthalmicformulation disclosed herein can comprise a non-ionic detergent. In someembodiments, the non-ionic detergent is a nonionic polymer containing apolyoxyethylene moiety. In some embodiments, the non-ionic detergent isany one or more of polysorbate 20, poloxamer 188 or polyethylene glycol3350. In some embodiments, the non-ionic detergent is polysorbate 20. Insome embodiments, the non-ionic detergent is polysorbate 80. In someembodiments, a pharmaceutical formulation or ophthalmic formulationdisclosed herein can contain about 0.01% to about 1% non-ionicdetergent. For example, a formulation of the present disclosure cancomprise about 0.0085%, about 0.01%, about 0.02%, about 0.03%, about0.04%, about 0.05%, about 0.06%, about 0.07%, about 0.08%, about 0.09%,about 0.1%, about 0.11%, about 0.12%, about 0.13%, about 0.14%, about0.15%, about 0.16%, about 0.17%, about 0.18%, about 0.19%, about 0.20%,about 0.21%, about 0.22%, about 0.23%, about 0.24%, about 0.25%, about0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about0.9%, about 1%, about 1.1%, about 1.15%, about 1.2%, about 1.25%, about1.3%, about 1.35%, about 1.4%, about 1.45%, about 1.5%, about 1.55%,about 1.6%, about 1.65%, about 1.7%, about 1.75%, about 1.8%, about1.85%, about 1.9%, about 1.95%, or about 2% polysorbate 20, polysorbate80 or poloxamer 188.

In some embodiments, a pharmaceutical formulation or ophthalmicformulation disclosed herein can comprise a tonicity agent. In someembodiments, the tonicity agent is sodium chloride or potassiumchloride. In some embodiments, the tonicity agent is sodium chloride. Insome embodiments, the sodium chloride is present at a concentration ofabout 5 mM to about 100 mM, about 10 mM to about 50 mM, or about 40 mM.

In some embodiments, a pharmaceutical formulation or ophthalmicformulation disclosed herein can comprise a stabilizer. In someembodiments, the stabilizer is a thermal stabilizer that can stabilizean antibody or compound disclosed herein under conditions of thermalstress. In some embodiments, the stabilizer maintains greater than about93% of the compound or antibody in a native conformation when thesolution containing the compound or antibody and the thermal stabilizeris kept at about 45° C. for up to about 28 days. In some embodiments,the stabilizer prevents aggregation of the compound or antibody and lessthan 4% of the compound or antibody is aggregated when the solutioncontaining the compound or antibody and the thermal stabilizer is keptat about 45° C. for up to about 28 days. In some embodiments, thestabilizer maintains greater than about 96% of the compound or antibodyin a native conformation when the solution containing the compound orantibody and the thermal stabilizer is kept at about 37° C. for up toabout 28 days. In some embodiments, the stabilizer prevents aggregationof the compound or antibody and less than about 2% of the compound orantibody is aggregated when the solution containing the compound orantibody and the thermal stabilizer is kept at about 37° C. for up toabout 28 days.

In some embodiments, the thermal stabilizer is a sugar or sugar alcohol,for example, sucrose, sorbitol, glycerol, trehalose, or mannitol, or anycombination thereof. In some embodiments, the stabilizer is a sugar. Insome embodiments, the sugar is sucrose, mannitol or trehalose. In someembodiments, the stabilizer is sucrose. In some embodiments, apharmaceutical formulation or ophthalmic formulation disclosed hereincan comprise about 1% to about 20% sugar or sugar alcohol, about 2% toabout 18% sugar or sugar alcohol, about 3% to about 15% sugar or sugaralcohol, about 4% to about 10% sugar or sugar alcohol, or about 5% sugaror sugar alcohol. For example, a pharmaceutical formulation orophthalmic formulation of the present disclosure can comprise about 4%,about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%,about 12%, about 13%, or about 14% sugar or sugar alcohol (e.g.,sucrose, trehalose or mannitol). In some embodiments, the stabilizer isat a concentration of from about 1% w/v to about 20% w/v. In someembodiments, the stabilizer is sucrose at a concentration of from about1% w/v to about 15% w/v, or from about 1% w/v to about 10% w/v. In someembodiments, the stabilizer is sucrose at a concentration of 5% w/v orabout 5% w/v. In some embodiments, the stabilizer is sucrose at aconcentration of 7.5% w/v or about 7.5% w/v. In some embodiments, thestabilizer is sucrose at a concentration of 10% w/v or about 10% w/v. Insome embodiments, the stabilizer is sucrose at a concentration of 12.5%w/v or about 12.5% w/v. In some embodiments, the stabilizer is sucroseat a concentration of 15% w/v or about 15% w/v. In some embodiments, thestabilizer is sucrose at a concentration of 20% w/v or about 20% w/v.

Administration of Pharmaceutical Compositions

A pharmaceutical composition disclosed herein can be administered in atherapeutically-effective amount by various forms and routes including,for example, oral, topical, parenteral, intravenous injection,intravenous infusion, subcutaneous injection, subcutaneous infusion,intramuscular injection, intramuscular infusion, intradermal injection,intradermal infusion, intraperitoneal injection, intraperitonealinfusion, intracerebral injection, intracerebral infusion, subarachnoidinjection, subarachnoid infusion, intraocular injection, intraspinalinjection, intrasternal injection, ophthalmic administration,endothelial administration, local administration, intranasaladministration, intrapulmonary administration, rectal administration,intraarterial administration, intrathecal administration, inhalation,intralesional administration, intradermal administration, epiduraladministration, absorption through epithelial or mucocutaneous linings(e.g., oral mucosa, rectal and intestinal mucosa), intracapsularadministration, subcapsular administration, intracardiac administration,transtracheal administration, subcuticular administration, subarachnoidadministration, subcapsular administration, intraspinal administration,or intrasternal administration.

A pharmaceutical composition can be administered in a local manner, forexample, via injection of the compound directly into an organ,optionally in a depot or sustained release formulation or implant. Apharmaceutical composition can be provided in the form of a rapidrelease formulation, in the form of an extended release formulation, orin the form of an intermediate release formulation. A rapid release formcan provide an immediate release. An extended release formulation canprovide a controlled release or a sustained delayed release.

In some embodiments, a pump can be used for delivery of thepharmaceutical composition. In some embodiments, a pen delivery devicecan be used, for example, for subcutaneous delivery of a composition ofthe disclosure. Such a pen delivery device can be reusable ordisposable. A reusable pen delivery device can use a replaceablecartridge that contains a pharmaceutical composition disclosed herein.Once all of the pharmaceutical composition within the cartridge has beenadministered and the cartridge is empty, the empty cartridge can readilybe discarded and replaced with a new cartridge that contains thepharmaceutical composition. The pen delivery device can then be reused.A disposable pen has no replaceable cartridge. Rather, the disposablepen delivery device comes prefilled with the pharmaceutical compositionheld in a reservoir within the device. Once the reservoir is emptied ofthe pharmaceutical composition, the entire device is discarded.

A pharmaceutical composition disclosed herein can be administered, forexample, to the eye via any suitable form or route including, forexample, topical, oral, systemic, intravitreal, intracameral,intracanieral, subconjunctival, subtenon, retrobulbar, intraocular,intrastromal, intracorneal, posterior juxtascleral, periocular,subretinal, or suprachoroidal administration. The delivery method caninclude an invasive method for direct delivery of the composition toocular cells. In some embodiments, a liquid pharmaceutical compositioncomprising an antibody or compound can be delivered via a subretinalinjection, intravitreal injection (e.g., front, mid or back vitrealinjection), intravitreal implant, intraorbital injection, intraorbitaladministration, subcutaneous injection, intracameral injection,intracanieral injection, subconjunctival injection, subconjunctivalimplant, injection into the anterior chamber via the temporal limbus,intrastromal injection, intracorneal injection, aqueous humor injection,subtenon injection, or subtenon implant. The compositions can beadministered by injecting the formulation in any part of the eyeincluding anterior chamber, posterior chamber, vitreous chamber(intravitreal), retina proper, and/or subretinal space.

A pharmaceutical composition disclosed herein can be delivered via anon-invasive method. Examples of non-invasive modes of administering theformulation can include using a needleless injection device, and topicaladministration, for example, eye drops to the cornea. Multipleadministration routes can be employed for efficient delivery of thepharmaceutical composition. In some embodiments, the composition isdelivered via multiple administration routes, for example, subretinaland intravitreous, to increase the efficiency of antibody delivery. Insome embodiments, the subretinal and/or intravitreal injection ispreceded by a vitrectomy.

In some embodiments, a liquid formulation comprising from 10 mg/mL to120 mg/mL of antibody or compound is in a prefilled syringe and isadministered intravitreally in a volume of up to about 500 μL. In someembodiments, a liquid formulation comprising from 10 mg/mL to 120 mg/mLof antibody or compound is in a prefilled syringe and is administeredintravitreally in a volume of up to about 100 μL. In some embodiments, aliquid formulation comprising from 10 mg/mL to 120 mg/mL of antibody orcompound is in a prefilled syringe and is administered intravitreally ina volume of about 50 μL.

A pharmaceutical composition disclosed herein can be targeted to anysuitable ocular cell including, for example, endothelial cells such asvascular endothelial cells, cells of the retina such as retinal pigmentepilthelium (RPE), corneal cells, fibroblasts, astrocytes, glial cells,pericytes, iris epithelial cells, cells of neural origin, ciliaryepithelial cells, Müller cells, muscle cells surrounding and attached tothe eye such as cells of the lateral rectus muscle, orbital fat cells,cells of the sclera and episclera, cells of the trabecular meshwork, orconnective tissue cells.

Dosing

A compound, antibody, or therapeutic agent described herein can beadministered before, during, or after the occurrence of a disease orcondition, and the timing of administering a composition containing thecompound, antibody, or therapeutic agent can vary. For example, thecomposition can be used as a prophylactic and can be administeredcontinuously to subjects with a propensity to conditions or diseases inorder to lessen a likelihood of the occurrence of the disease orcondition. The composition can be administered to a subject alreadysuffering from a disease or condition, in an amount sufficient to cureor at least partially arrest the symptoms of the disease or condition,or to cure, heal, improve, or ameliorate the condition. The compositioncan be administered to a subject during or as soon as possible after theonset of the symptoms. The administration of the compound, antibody, ortherapeutic agent can be initiated within the first 48 hours of theonset of the symptoms, within the first 24 hours of the onset of thesymptoms, within the first 6 hours of the onset of the symptoms, orwithin 3 hours of the onset of the symptoms. The initial administrationcan be via any practical route, such as by any route described hereinusing any formulation described herein. The compound, antibody, ortherapeutic agent can be administered as soon as is practicable afterthe onset of a disease or condition is detected or suspected, and for alength of time necessary for the treatment of the disease, such as, forexample, from about 1 month to about 3 months. The length of treatmentcan vary for each subject. Amounts effective for this use can vary basedon the severity and course of the disease or condition, previoustherapy, the subject's health status, weight, and response to the drugs,and the judgment of the treating physician. Improvement of clinicalsymptoms can be monitored, for example, by indirect ophthalmoscopy,fundus photography, fluorescein angiography, electroretinography,external eye examination, slit lamp biomicroscopy, applanationtonometry, pachymetry, optical coherence tomography, or autorefraction.

A pharmaceutical composition described herein can be in a unit dosageform suitable for a single administration of a precise dosage. In unitdosage form, the formulation can be divided into unit doses containingappropriate quantities of one or more compounds, antibodies ortherapeutic agents. The unit dosage can be in the form of a packagecontaining discrete quantities of the formulation. Non-limiting examplesare packaged injectables, vials, and ampoules. An aqueous suspensioncomposition disclosed herein can be packaged in a single-dosenon-reclosable container. Multiple-dose reclosable containers can beused, for example, in combination with or without a preservative. Aformulation for injection disclosed herein can be present in a unitdosage form, for example, in ampoules, or in multi-dose containers witha preservative.

Multiple compounds, antibodies or therapeutic agents disclosed hereincan be administered in any order or simultaneously. If simultaneously,the multiple compounds, antibodies or therapeutic agents can be providedin a single, unified form, or in multiple forms, for example, asmultiple separate injections or infusions. The compounds, antibodies ortherapeutic agents can be packed together or separately, in a singlepackage or in a plurality of packages. One or all of the compounds,antibodies or therapeutic agents can be given in multiple doses. If notsimultaneous, the timing between the multiple doses can vary to as muchas about a month.

An intraocular injection can be performed between any interval of timeto improve efficiency of delivery and/or to minimize or avoid damage tosurrounding tissue. The interval of time between two or more intraocularinjections can be from, for example, about 1 minute to about 60 minutes,about 1 minute to about 5 minutes, about 5 minutes to about 10 minutes,about 10 minutes to about 15 minutes, about 15 minutes to about 20minutes, about 20 minutes to about 25 minutes, about 25 minutes to about30 minutes, about 30 minutes to about 35 minutes, about 35 minutes toabout 40 minutes, about 40 minutes to about 45 minutes, about 45 minutesto about 50 minutes, about 50 minutes to about 55 minutes, or about 55minutes to about 60 minutes.

An intraocular injection can be performed at any rate. The rate ofintraocular injection can be from, for example, about 1 μL/sec to about500 μL/sec, about 1 μL/sec to about 10 μL/sec, about 10 μL/sec to about20 μL/sec, about 20 μL/sec to about 30 μL/sec, about 30 μL/sec to about40 μL/sec, about 40 μL/sec to about 50 μL/sec, about 50 μL/sec to about60 μL/sec, about 60 μL/sec to about 70 μL/sec, about 70 μL/sec to about80 μL/sec, about 80 μL/sec to about 90 L/sec, about 90 μL/sec to about100 μL/sec, about 100 μL/sec to about 110 μL/sec, about 110 L/sec toabout 120 μL/sec, about 120 μL/sec to about 130 μL/sec, about 130 μL/secto about 140 μL/sec, about 140 μL/sec to about 150 μL/sec, about 150μL/sec to about 160 μL/sec, about 160 μL/sec to about 170 μL/sec, about170 μL/sec to about 180 μL/sec, about 180 μL/sec to about 190 μL/sec,about 190 μL/sec to about 200 μL/sec, about 200 μL/sec to about 300μL/sec, about 300 μL/sec to about 400 μL/sec, or about 400 μL/sec toabout 500 μL/sec.

A compound, antibody, or therapeutic agent disclosed herein can beadministered at a dosage of about 0.0001 mg/kg to about 1000 mg/kg,about 0.001 mg/kg to about 100 mg/kg, about 0.01 mg/kg to about 100mg/kg, about 0.01 mg/kg to about 20 mg/kg, about 0.02 mg/kg to about 7mg/kg, about 0.03 mg/kg to about 5 mg/kg, about 0.05 mg/kg to about 3mg/kg, about 0.1 mg/kg to about 50 mg/kg, about 0.1 mg/kg to about 0.5mg/kg, about 0.2 mg/kg to about 0.6 mg/kg, about 0.3 mg/kg to about 0.7mg/kg, about 0.4 mg/kg to about 0.8 mg/kg, about 0.1 mg/kg to about 0.9mg/kg, about 0.01 mg/kg to about 50 mg/kg, about 0.1 mg/kg to about 10mg/kg, about 1 mg/kg to about 10 mg/kg, about 5 mg/kg to about 10 mg/kg,about 1 mg/kg to about 5 mg/kg, or about 3 mg/kg to about 7 mg/kg bymass of the subject.

A compound, antibody, or therapeutic agent described herein can beadministered at any interval desired. The administration of thecompound, antibody, or therapeutic agent can have regular or irregulardosing schedules to accommodate either the person administering thecompound, antibody, or therapeutic agent or the subject receiving thecompound, antibody, or therapeutic agent. For example, the compound,antibody, or therapeutic agent can be administered twice a day, once aday, five times a week, four times a week, three times a week, two timesa week, once a week, once every two weeks, once every three weeks, onceevery four weeks, once a month, once every five weeks, once every sixweeks, once every eight weeks, once every two months, once every twelveweeks, once every three months, once every four months, once every sixmonths, once a year, or less frequently. In some embodiments,administration is every other week.

The amount administered can be of the same amount in each dose or thedosage can vary between doses. For example, a first amount can beadministered in the morning and a second amount can be administered inthe evening.

A compound, antibody, or therapeutic agent described herein can beadministered in any amount necessary or convenient. For example, acompound described herein can be administered in an amount from about0.05 mg to about 300 mg, about 0.1 mg to about 300 mg, about 0.1 mg toabout 200 mg, about 0.1 mg to about 100 mg, about 0.05 mg to about 1.5mg, about 0.1 mg to about 1.5 mg, about 0.05 mg to about 1 mg, about 1mg to about 1.5 mg, about 0.5 mg to about 6 mg, about 1 mg to about 4mg, about 2 mg to about 10 mg, about 10 mg to about 30 mg, about 30 mgto about 50 mg, about 50 mg to about 70 mg, about 70 mg to about 100 mg,or about 0.1 mg to about 1 mg, about 0.05 mg, about 0.06 mg, about 0.07mg, about 0.08 mg, about 0.09 mg, about 0.1 mg, about 0.11 mg, about0.12 mg, about 0.13 mg, about 0.14 mg, about 0.15 mg, about 0.16 mg,about 0.17, mg, about 0.18 mg, about 0.19 mg, about 0.2 mg, about 0.21mg, about 0.22 mg, about 0.23 mg, about 0.24 mg, about 0.25 mg, about0.26 mg, about 0.27, mg, about 0.28 mg, about 0.29 mg, about 0.3 mg,about 0.31 mg, about 0.32 mg, about 0.33 mg, about 0.34 mg, about 0.35mg, about 0.36 mg, about 0.37, mg, about 0.38 mg, about 0.39 mg, about0.4 mg, about 0.41 mg, about 0.42 mg, about 0.43 mg, about 0.44 mg,about 0.45 mg, about 0.46 mg, about 0.47, mg, about 0.48 mg, about 0.49mg, about 0.5 mg, about 0.51 mg, about 0.52 mg, about 0.53 mg, about0.54 mg, about 0.55 mg, about 0.56 mg, about 0.57, mg, about 0.58 mg,about 0.59 mg, about 0.6 mg, about 0.61 mg, about 0.62 mg, about 0.63mg, about 0.64 mg, about 0.65 mg, about 0.66 mg, about 0.67, mg, about0.68 mg, about 0.69 mg, about 0.7 mg, about 0.71 mg, about 0.72 mg,about 0.73 mg, about 0.74 mg, about 0.75 mg, about 0.76 mg, about 0.77,mg, about 0.78 mg, about 0.79 mg, about 0.8 mg, about 0.81 mg, about0.82 mg, about 0.83 mg, about 0.84 mg, about 0.85 mg, about 0.86 mg,about 0.87, mg, about 0.88 mg, about 0.89 mg, about 0.9 mg, about 0.91mg, about 0.92 mg, about 0.93 mg, about 0.94 mg, about 0.95 mg, about0.96 mg, about 0.97, mg, about 0.98 mg, about 0.99 mg, about 1 mg, about1.5 mg, about 2 mg, about 2.5 mg, about 3 mg, about 3.5 mg, about 4 mg,about 4.5 mg, about 5 mg, about 5.5 mg, about 6 mg, about 6.5 mg, about7 mg, about 7.5 mg, about 8 mg, about 8.5 mg, about 9 mg, about 9.5 mg,about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg,about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about90 mg, about 95 mg, about 100 mg, about 110 mg, about 120 mg, about 130mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180mg, about 190 mg about 200 mg, about 210 mg, about 220 mg, about 230 mg,about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg,about 290 mg, or about 300 mg per dose for a subject by any route ofadministration.

Combination Therapies

A pharmaceutical composition provided herein can be administered inconjunction with other therapies, for example, chemotherapy, radiation,surgery, anti-inflammatory agents, or vitamins. The other agents can beadministered prior to, after, or concomitantly with the pharmaceuticalcomposition.

In some embodiments, a compound or antibody described herein can be usedsingly or in combination with one or more therapeutic agents as acomponent of mixtures.

In some embodiments, the disclosure provides co-administration of amultispecific compound or antibody targeting, for example, HPTP-β(VE-PTP) and VEGF, with one or more additional anti-VEGF agents, whichcan stabilize the vasculature against neovascularization. In someembodiments, co-administration of the multispecific compound or antibodywith one or more additional anti-VEGF agents can stabilize thevasculature against leakage. An anti-VEGF agent can be a compound, arecombinant protein, an antibody, an antigen-binding fragment, variant,or derivative thereof (e.g., a scFv), a protein comprising one or moredesigned ankyrin repeats, a designed ankyrin repeat protein (DARPin), anankyrin protein, an ankyrin repeat protein, an affibody, an avimer, anadnectin, an anticalin, a Fynomer, a Kunitz domain, a knottin, aβ-hairpin mimetic, or a peptide derived from one or more receptors, e.g.VEGF receptors, or the VEGF-binding portions of human VEGF receptors 1and 2.

Non-limiting examples of anti-VEGF agents include bevacizumab(Avastin®), ranibizumab (Lucentis®), aflibercept (Eylea®), conbercept,brolucizumab, RTH258, VEGF receptor tyrosine kinase inhibitors such assorafenib, sunitinib, axitinib, pazopanib, vandetinib, cabozantinib,regorafenib, and4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline(ZD6474), VEGF variants, soluble VEGF receptor fragments or traps,aptamers capable of blocking VEGF or VEGFR (e.g. Pegaptanib),neutralizing anti-VEGFR antibodies or fragments thereof (e.g.,ramucirumab, p1C11, 1121, 1121B), anti-KDR antibodies, anti-fit1antibodies, low molecular weight inhibitors of VEGFR tyrosine kinases,DARPins that bind VEGF (e.g., abicipar, MP0112, MP0250), proteinscomprising one or more designed ankyrin repeats that bind VEGF,adnectins (e.g., CT-322), anticalins (e.g., PRS-050), collagenIV-derived biomimetic peptides (e.g., AXT-107)

Further non-limiting examples of VEGF-modulating agents includeanti-inflammatory agents, for example, dexamethasone, fluocinolone, andtriamcinolone. The multispecific compound or antibody targeting, forexample, HPTP-β (VE-PTP) and VEGF, can be administered in combinationwith any additional anti-VEGF agent in any combination, for example, atthe beginning of treatment, at any time during treatment, or at any timeafter treatment with the additional anti-VEGF agent has concluded. Inaddition, the dosage of the multispecific compound or antibody can beadjusted during treatment. Also, the dosage of the additional anti-VEGFagent can be adjusted during treatment. The multispecific compound orantibody can be administered, for example, monthly, once every 3 months,once every 6 months, or yearly, wherein the additional anti-VEGF agentis administered at any frequency between treatments. Also disclosedherein are methods for treating a disease or condition as disclosedherein. The method comprises administering to a subject:

-   -   a) a therapeutically-effective amount of a multispecific        compound or antibody targeting, for example, HPTP-β (VE-PTP) and        VEGF; and    -   b) a therapeutically-effective amount of an additional anti-VEGF        agent;        wherein the administration of the multispecific compound or        antibody and the additional anti-VEGF agent can be conducted as        described herein.

In some embodiments, the disclosure provides co-administration of amultispecific compound or antibody targeting, for example, HPTP-β(VE-PTP) and VEGF, with one or more additional anti-HPTP-β (VE-PTP)agents, which can stabilize the vasculature against neovascularization.In some embodiments, co-administration of the multispecific compound orantibody with one or more additional anti-HPTP-β (VE-PTP) agents canstabilize the vasculature against leakage. An anti-HPTP-β (VE-PTP) agentcan be a compound, a recombinant protein, an antibody, anantigen-binding fragment, variant, or derivative thereof (e.g., a scFv),a protein comprising one or more designed ankyrin repeats, a designedankyrin repeat protein (DARPin), an ankyrin protein, an ankyrin repeatprotein, an affibody, an avimer, an adnectin, an anticalin, a Fynomer, aKunitz domain, a knottin, a β-hairpin mimetic, or a peptide derived fromone or more receptors. In some embodiments, the additional anti-HPTP-β(VE-PTP) agent(s) can activate Tie2 signaling by promoting proteinphosphorylation, such as phosphorylation of the Tie2 protein. In someembodiments, the additional anti-HPTP-β (VE-PTP) agent(s) can bind toHPTP-β (VE-PTP).

The multispecific compound or antibody targeting, for example, HPTP-β(VE-PTP) and VEGF, can be administered in combination with anyadditional anti-HPTP-β (VE-PTP) agent in any combination, for example,at the beginning of treatment, at any time during treatment, or at anytime after treatment with the additional anti-HPTP-β (VE-PTP) agent hasconcluded. The dosage of the multispecific compound or antibody can beadjusted during treatment. The dosage of the additional anti-HPTP-β(VE-PTP) agent can be adjusted during treatment. The multispecificcompound or antibody can be administered, for example, monthly, onceevery 3 months, once every 6 months, or yearly, wherein the additionalanti-HPTP-β (VE-PTP) agent is administered at any frequency betweentreatments. Also disclosed herein are methods for treating a disease orcondition as disclosed herein. The method comprises administering to asubject:

-   -   a) a therapeutically-effective amount of a multispecific        compound or antibody targeting, for example, HPTP-β (VE-PTP) and        VEGF; and    -   b) a therapeutically-effective amount of an additional        anti-HPTP-β (VE-PTP) agent;        wherein the administration of the multispecific compound or        antibody and the additional anti-HPTP-β (VE-PTP) agent can be        conducted as described herein.

In some embodiments, the disclosure provides co-administration of amultispecific compound or antibody targeting, for example, HPTP-β(VE-PTP) and VEGF, with one or more additional Tie2 receptor activatingcompounds. An additional Tie2 receptor activating compound can be, forexample, an angiopoietin 1 recombinant protein, an Ang1 mimetic, a Tie2agonist, a peptide, a HPTP-β (VE-PTP) phosphatase inhibitor, aTie2-peptomimetic, a tetrameric polyethylene oxide clustered peptide, acollagen IV-biomimetic peptide, a compound, a recombinant protein, anantibody, an antigen-binding fragment, variant, or derivative thereof(e.g., an scFv), an affinibody, an avimer, an adnectin, a proteincomprising one or more designed ankyrin repeats, a designed ankyrinrepeat protein (DARPin), an ankyrin protein, an ankyrin repeat protein,an affibody, an avimer, an adnectin, an anticalin, a Fynomer, a Kunitzdomain, a knottin, a β-hairpin mimetic, or a peptide derived from one ormore receptors. In some embodiments, the one or more additional Tie2receptor activating compounds are small molecules. In some embodiments,the one or more additional Tie2 receptor activating compounds improvedrainage through ocular lymphatics, Schlemm's canal, or corneal limballymphatics. In some embodiments, the one or more additional Tie2receptor activating compounds are administered as eye drops. In someembodiments, the one or more additional Tie2 receptor activatingcompounds are administered to treat primary open angle glaucoma,age-related macular degeneration, cardiovascular disease, or cystickidney disease. In some embodiments, the one or more additional Tie2receptor activating compounds can be, for example, MAN-01, AXT-107, orvasculotide. In some embodiments, a compound disclosed herein can beco-administered with, for example, MAN-01, AXT-107, or vasculotide.

The multispecific compound or antibody targeting, for example, HPTP-β(VE-PTP) and VEGF, can be administered in combination with anyadditional Tie2 receptor activating compound in any combination, forexample, at the beginning of treatment, at any time during treatment, orat any time after treatment with the additional Tie2 receptor activatingcompound has concluded. The dosage of the multispecific compound orantibody can be adjusted during treatment. The dosage of the additionalTie2 receptor activating compound can be adjusted during treatment. Themultispecific compound or antibody can be administered, for example,monthly, once every 3 months, once every 6 months, or yearly, whereinthe additional Tie2 receptor activating compound is administered at anyfrequency between treatments with the multispecific compound orantibody. Also disclosed herein are methods for treating a disease orcondition as disclosed herein. The method comprises administering to asubject:

-   -   a) a therapeutically-effective amount of a multispecific        compound or antibody targeting, for example, HPTP-β (VE-PTP) and        VEGF; and    -   b) a therapeutically-effective amount of a Tie2 activator;        wherein the administration of the multispecific compound or        antibody and the additional Tie2 receptor activating compound        can be conducted as described herein.        Mouse Models        Oxygen-Induced Ischemic Retinopathy Model

The oxygen-induced ischemic retinopathy model can be considered to mimicaspects of proliferative retinal neovascularization and proliferativediabetic retinopathy. One week old mice can be placed in an airtightchamber and exposed to hyperoxia (75±3% oxygen) for five days, resultingin hyperoxia-induced neovascularization, for example, at the junctionbetween the vascularized and avascular retina between postnatal day 17and 21. Mice can be dosed with a compound of interest, for example, anantibody or compound of the disclosure, to determine the effect onneovascularization and/or vascular leakage. Neovascularization and/orvascular leakage can be assessed as described below.

Rho/VEGF Mouse Model

The Rho/VEGF mouse model can mimic aspects of neovascular age-relatedmacular degeneration. Transgenic mice with vascular endothelial growthfactor (VEGF) expression driven by the rhodopsin promoter (rho/VEGFmice) can develop retinal neovascularization, retinal angiomatousproliferation, and retinal vascular leakage. In rho/VEGF mice, VEGFexpression in photoreceptors can begin between postnatal days 5 and 10,the period when the deep capillary bed is developing. Neovascularizationcan originate from the deep capillary bed of the retina and grow intothe subretinal space. Mice can be dosed with a compound of interest, forexample, an antibody or compound of the disclosure, to determine theeffect on neovascularization and/or vascular leakage. Neovascularizationand/or vascular leakage can be assessed as described below.

Tet/Opsin/VEGF Mouse Model

Mice with a VEGF under the control of a reverse tetracyclinetransactivator (rtTA) inducible promoter coupled to the rhodopsinpromoter (Tet/opsin/VEGF mice) can be used as an inducible model ofneovascularization, retinal vascular leakage, and retinal detachment. Inthese mice, VEGF transgene expression in the retina can be induced byadministering doxycycline. Neovascularization can be evident by three tofour days after VEGF induction. Neovascularization can be moreextensive, and can cause outer retinal folds followed by total retinaldetachment within about five days. Mice can be dosed with a compound ofinterest, for example, an antibody or compound of the disclosure, todetermine the effect on neovascularization, vascular leakage, and/orretinal detachment. Neovascularization and/or vascular leakage can beassessed as described below. To assess retinal detachment, eyes can befrozen in cutting temperature embedding solution, ten micron sectionscut through the entire eye, and sections stained with Hoechst. Sectionscan be examined by light microscopy, the mean length of the retinaldetachment per section measured by image analysis, and the detachedpercentage of the retina calculated.

Tet/Opsin/Ang2 Mouse Model

Tet/opsin/Ang2 mice have inducible expression of Ang2 in the retina.These mice can be used to study the impact of Ang2 expression in variousexperimental conditions. For example, Tet/opsin/Ang2 mice can be used todetermine the effect of Ang2 expression on neovascularization when VEGFlevels are high versus low, or the effect of Ang2 expression in theoxygen induced ischemic retinopathy model. Mice can be dosed with acompound of interest, for example, an antibody or compound of thedisclosure, to determine the impact of Ang2 expression on therapeuticefficacy.

Laser-Induced Choroidal Neovascularization Model

The laser-induced choroidal neovascularization model can be consideredto mimic aspects of neovascular age-related macular degeneration.Anesthetized mice can have their pupils dilated, and burns can bedelivered, for example, to the retina by a krypton laser using a slitlamp system and a cover glass as a contact lens. Multiple burns can beproduced in a single eye, for example, burns in three locations per eye.Burns can cause rupture of Bruch's membrane. Choroidalneovascularization can be assessed at various time points after lasertreatment, for example, one week, two weeks, or four weeks after lasertreatment. Mice can be dosed with a compound of interest, for example,an antibody or compound of the disclosure, to determine the effect onneovascularization. Eyecups can be stained with FITC-labeled GSA,choroids flat mounted, and the area of choroidal neovascularization ateach Bruch's membrane rupture site measured with fluorescence microscopyand image analysis. Neovascularization and/or vascular leakage can alsobe assessed as described below.

Assessment of Neovascularization

Neovascularization can be assessed using a range of techniques.

Fluorescein angiograms can be done by taking serial fundus photographsafter injection of a dye to reveal blood vessels, allowingidentification of the presence, location, and size of a neovascularcomplex. For example, an intraperitoneal injection of 0.3 mL of 1%fluorescein sodium can be given, serial fundus photographs taken, andthe choroidal neovascularization area, total lesion area, and leakagearea can be measured.

Eyes can be processed for evaluation by fluorescent, light, or electronmicroscopy. For example, mice can be perfused with fluorescentlylabelled dextran, or eyes can be injected with GSA or antibodiestargeting PECAM. Eyes can be processed for observation under afluorescent microscope, and the extent of neovascularization can bequantified, for example, by quantifying the area of neovascularizationper retina, by quantifying the area of neovascularization at eachBruch's membrane rupture site, or by quantifying the number of nuclei ofnew vessels extending from the retina into the vitreous.

The area of retinal neovascularization can be determined, for example,using FITC-labeled GSA lectin and fluorescence microscopy. Eyes can befixed in 10% formalin, dissected intact, washed with PBS, blocked in 8%swine serum, and stained with FITC-labeled GSA lectin for a timeappropriate to stain retinal neovascularization and hyaloid vessels, butnot normal retinal vessels (e.g., 40-50 minutes). Retinas can be flatmounted, digital images can be obtained with a fluorescent microscopeand merged into a single image of the entire retina. Software can beused to measure the area of neovascularization per retina.

The area of subretinal neovascularization can be determined, forexample, using FITC-labeled GSA lectin and fluorescence microscopy. Eyescan be fixed in 10% formalin, and retinas can be dissected, blocked with5% swine serum, stained with FITC-conjugated GSA for two hours to stainvascular cells, and flat mounted with the photoreceptor side up. Thearea of subretinal neovascularization can be measured with fluorescencemicroscopy and image analysis.

Assessment of Retinal Vascular Leakage

Retinal vascular leakage can be assessed by measuring extravasated serumalbumin using an immunofluorescent technique. For example, eyes can befixed in 10% formalin, retinas can be dissected, washed, blocked, andstained using an anti-albumin antibody and a fluorescently-conjugatedsecondary antibody. The vessels can be labeled by counterstaining withGSA lectin, and retinas flat mounted. Retinas can be examined byfluorescence microscopy, and the area of albumin staining determined byimage analysis. Retinal vascular leakage is relevant to, for example,diabetic macular edema and macular edema due to retinal vein occlusion.

Miles Assay for Vascular Leakage

The Miles assay can be used to assess vascular leakage in dermalsubcutaneous blood vessels. Evans blue is a dye that binds albumin.Under physiologic conditions the endothelium is impermeable to albumin,so Evans blue bound albumin remains restricted within blood vessels. Inpathologic conditions that promote increased vascular permeability,endothelial cells partially lose close contacts. The endothelium becomespermeable to small proteins such as albumin. Mice can be injectedintravenously with 1% Evans blue dye in PBS, and injected intradermallywith VEGF. Thirty minutes after intradermal injections, tissue at theintradermal injection sites can be excised and extracted in formamide toassess Evans blue dye extravasation. Vascular leakage can be quantifiedby measurement of the dye incorporated per milligram of tissue, forexample, using optical density measurements and a standard curve. Micecan be dosed with a compound of interest, for example, an antibody orcompound of the disclosure, to determine the effect on vascular leakage.

Cancer Models

The efficacy of a compound or antibody of the disclosure as a treatmentfor cancer can be tested in a range of cancer models. In some cancermodels, cancer cells from a cell line can be implanted in a recipientanimal, for example, a mouse. Non-limiting examples of suitable mousestrains include C57BL6, BALB/C, C3H, FVB/N, andFVB/N-Tg(IMTV-PyVT)634Mul. Non-limiting examples of suitable cancer celllines include 4T1, E0771, and P0008. Cells of the 4T1 or E0771 celllines can, for example, be implanted into a mammary pad as a model ofbreast cancer. 4T1 or E0771 cells can be implanted, for example, intothe third mammary fat pad of female nude mice.

The size of solid tumors can be measured with calipers and tumor volumecalculated. Tumors can be processed for histopathological evaluation orfluorescence microscopy, to assess, for example, tumor area, tumorgrade, metastases count, metastases area, the number of cell nuclei pertumor focus, intratumoral vessel diameter, intratumoral vessel density,tumor vascular maturity, perivascular cell coverage, proximity betweenperivascular cells and endothelial cells, or to grade tumors foci asintravascular or extravascular.

Spontaneous metastasis models can be used to study the effects of acompound or antibody of the disclosure on metastasis. After tumorimplant, a primary tumor can be resected (e.g., upon reaching 5 mm insize), and the animal later evaluated macroscopically orhistopathologically for metastases, for example, to determine the numberand size of metastases in the lungs, liver, lymph nodes, and bones.

To evaluate the impact of vessel stability on metastasis, a model can beused wherein tumor cells can be injected intravenously, and the animalsubsequently evaluated for intravascular versus extravasated metastases.For example, 4T-1 cells can be injected intravenously, and the lungsprocessed for histopathologic evaluation to quantify intravascularversus extravascular metastases.

Metastases can be counted and measured macroscopically, for example, byimmersing lungs in Bouin's solution, then examining them with astereomicroscope.

To measure tumor vessel permeability in vivo, intravital multiphotonmicroscopy can be used. Animals can be injected with fluorescentlylabeled bovine serum albumin, for example, pre-treatment andpost-treatment time points. At each time point, two distinct regionswithin the tumor can be selected and a 200 micron image stack recordingBSA fluorescence taken through each region every ten minutes for anhour, using a multiphoton laser scanning microscope. The analysisapproach can involve three-dimensional vessel tracing to create vesselmetrics and a three-dimensional map of voxel intensity versus distanceto the nearest vessel over time. Images can be corrected for samplemovement over time with three-dimensional image registration. Thenormalized transvascular flux can be calculated.

Illustrative CDR Combinations

In some embodiments, an antibody or compound of this disclosurecomprises a HCDR1. In some embodiments, an antibody or compound of thisdisclosure comprises a HCDR2. In some embodiments, an antibody orcompound of this disclosure comprises a HCDR3. In some embodiments, anantibody or compound of this disclosure comprises a LCDR1. In someembodiments, an antibody or compound of this disclosure comprises aLCDR2. In some embodiments, an antibody or compound of this disclosurecomprises a LCDR3.

In some embodiments, an antibody or compound of this disclosurecomprises a HCDR1 and a HCDR2. In some embodiments, an antibody orcompound of this disclosure comprises a HCDR1 and a HCDR3. In someembodiments, an antibody or compound of this disclosure comprises aHCDR1 and a LCDR1. In some embodiments, an antibody or compound of thisdisclosure comprises a HCDR1 and a LCDR2. In some embodiments, anantibody or compound of this disclosure comprises a HCDR1 and a LCDR3.In some embodiments, an antibody or compound of this disclosurecomprises a HCDR2 and a HCDR3. In some embodiments, an antibody orcompound of this disclosure comprises a HCDR2 and a LCDR1. In someembodiments, an antibody or compound of this disclosure comprises aHCDR2 and a LCDR2. In some embodiments, an antibody or compound of thisdisclosure comprises a HCDR2 and a LCDR3. In some embodiments, anantibody or compound of this disclosure comprises a HCDR3 and a LCDR1.In some embodiments, an antibody or compound of this disclosurecomprises a HCDR3 and a LCDR2. In some embodiments, an antibody orcompound of this disclosure comprises a HCDR3 and a LCDR3. In someembodiments, an antibody or compound of this disclosure comprises aLCDR1 and a LCDR2. In some embodiments, an antibody or compound of thisdisclosure comprises a LCDR1 and a LCDR3. In some embodiments, anantibody or compound of this disclosure comprises a LCDR2 and a LCDR3.

In some embodiments, an antibody or compound of this disclosurecomprises a HCDR1, a HCDR2, and a HCDR3. In some embodiments, anantibody or compound of this disclosure comprises a HCDR1, a HCDR2, anda LCDR1. In some embodiments, an antibody or compound of this disclosurecomprises a HCDR1, a HCDR2, and a LCDR2. In some embodiments, anantibody or compound of this disclosure comprises a HCDR1, a HCDR2, anda LCDR3. In some embodiments, an antibody or compound of this disclosurecomprises a HCDR1, a HCDR3, and a LCDR1. In some embodiments, anantibody or compound of this disclosure comprises a HCDR1, a HCDR3, anda LCDR2. In some embodiments, an antibody or compound of this disclosurecomprises a HCDR1, a HCDR3, and a LCDR3. In some embodiments, anantibody or compound of this disclosure comprises a HCDR1, a LCDR1, anda LCDR2. In some embodiments, an antibody or compound of this disclosurecomprises a HCDR1, a LCDR1, and a LCDR3. In some embodiments, anantibody or compound of this disclosure comprises a HCDR1, a LCDR2, anda LCDR3. In some embodiments, an antibody or compound of this disclosurecomprises a HCDR2, a HCDR3, and a LCDR1. In some embodiments, anantibody or compound of this disclosure comprises a HCDR2, a HCDR3, anda LCDR2. In some embodiments, an antibody or compound of this disclosurecomprises a HCDR2, a HCDR3, and a LCDR3. In some embodiments, anantibody or compound of this disclosure comprises a HCDR2, a LCDR1, anda LCDR2. In some embodiments, an antibody or compound of this disclosurecomprises a HCDR2, a LCDR1, and a LCDR3. In some embodiments, anantibody or compound of this disclosure comprises a HCDR2, a LCDR2, anda LCDR3. In some embodiments, an antibody or compound of this disclosurecomprises a HCDR3, a LCDR1, and a LCDR2. In some embodiments, anantibody or compound of this disclosure comprises a HCDR3, a LCDR1, anda LCDR3. In some embodiments, an antibody or compound of this disclosurecomprises a HCDR3, a LCDR2, and a LCDR3. In some embodiments, anantibody or compound of this disclosure comprises a LCDR1, a LCDR2, anda LCDR3.

In some embodiments, an antibody or compound of this disclosurecomprises a HCDR1, a HCDR2, a HCDR3, and a LCDR1. In some embodiments,an antibody or compound of this disclosure comprises a HCDR1, a HCDR2, aHCDR3, and a LCDR2. In some embodiments, an antibody or compound of thisdisclosure comprises a HCDR1, a HCDR2, a HCDR3, and a LCDR3. In someembodiments, an antibody or compound of this disclosure comprises aHCDR1, a HCDR2, a LCDR1, and a LCDR2. In some embodiments, an antibodyor compound of this disclosure comprises a HCDR1, a HCDR2, a LCDR1, anda LCDR3. In some embodiments, an antibody or compound of this disclosurecomprises a HCDR1, a HCDR2, a LCDR2, and a LCDR3. In some embodiments,an antibody or compound of this disclosure comprises a HCDR1, a HCDR3, aLCDR1, and a LCDR2. In some embodiments, an antibody or compound of thisdisclosure comprises a HCDR1, a HCDR3, a LCDR1, and a LCDR3. In someembodiments, an antibody or compound of this disclosure comprises aHCDR1, a HCDR3, a LCDR2, and a LCDR3. In some embodiments, an antibodyor compound of this disclosure comprises a HCDR1, a LCDR1, a LCDR2, anda LCDR3. In some embodiments, an antibody or compound of this disclosurecomprises a HCDR2, a HCDR3, a LCDR1, and a LCDR2. In some embodiments,an antibody or compound of this disclosure comprises a HCDR2, a HCDR3, aLCDR1, and a LCDR3. In some embodiments, an antibody or compound of thisdisclosure comprises a HCDR2, a HCDR3, a LCDR2, and a LCDR3. In someembodiments, an antibody or compound of this disclosure comprises aHCDR2, a LCDR1, a LCDR2, and a LCDR3. In some embodiments, an antibodyor compound of this disclosure comprises a HCDR3, a LCDR1, a LCDR2, anda LCDR3.

In some embodiments, an antibody or compound of this disclosurecomprises a HCDR1, a HCDR2, a HCDR3, a LCDR1, and a LCDR2. In someembodiments, an antibody or compound of this disclosure comprises aHCDR1, a HCDR2, a HCDR3, a LCDR1, and a LCDR3. In some embodiments, anantibody or compound of this disclosure comprises a HCDR1, a HCDR2, aHCDR3, a LCDR2, and a LCDR3. In some embodiments, an antibody orcompound of this disclosure comprises a HCDR1, a HCDR2, a LCDR1, aLCDR2, and a LCDR3. In some embodiments, an antibody or compound of thisdisclosure comprises a HCDR1, a HCDR3, a LCDR1, a LCDR2, and a LCDR3. Insome embodiments, an antibody or compound of this disclosure comprises aHCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3.

In some embodiments, an antibody or compound of this disclosurecomprises a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3.

In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 76-80. In some embodiments, an antibodyor compound of this disclosure comprises any one of SEQ ID NOS: 81-87.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 88-90. In some embodiments, an antibodyor compound of this disclosure comprises any one of SEQ ID NOS: 91-93.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 94-96. In some embodiments, an antibodyor compound of this disclosure comprises any one of SEQ ID NOS: 97-98.

In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 76-80 and any one of SEQ ID NOS: 81-87.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 76-80 and any one of SEQ ID NOS: 88-90.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 76-80 and any one of SEQ ID NOS: 91-93.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 76-80 and any one of SEQ ID NOS: 94-96.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 76-80 and any one of SEQ ID NOS: 97-98.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 81-87 and any one of SEQ ID NOS: 88-90.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 81-87 and any one of SEQ ID NOS: 91-93.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 81-87 and any one of SEQ ID NOS: 94-96.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 81-87 and any one of SEQ ID NOS: 97-98.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 88-90 and any one of SEQ ID NOS: 91-93.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 88-90 and any one of SEQ ID NOS: 94-96.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 88-90 and any one of SEQ ID NOS: 97-98.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 91-93 and any one of SEQ ID NOS: 94-96.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 91-93 and any one of SEQ ID NOS: 97-98.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 94-96 and any one of SEQ ID NOS: 97-98.

In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 76-80, any one of SEQ ID NOS: 81-87,and any one of SEQ ID NOS: 88-90. In some embodiments, an antibody orcompound of this disclosure comprises any one of SEQ ID NOS: 76-80, anyone of SEQ ID NOS: 81-87, and any one of SEQ ID NOS: 91-93. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 76-80, any one of SEQ ID NOS: 81-87, and any one ofSEQ ID NOS: 94-96. In some embodiments, an antibody or compound of thisdisclosure comprises any one of SEQ ID NOS: 76-80, any one of SEQ IDNOS: 81-87, and any one of SEQ ID NOS: 97-98. In some embodiments, anantibody or compound of this disclosure comprises any one of SEQ ID NOS:76-80, any one of SEQ ID NOS: 88-90, and any one of SEQ ID NOS: 91-93.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 76-80, any one of SEQ ID NOS: 88-90,and any one of SEQ ID NOS: 94-96. In some embodiments, an antibody orcompound of this disclosure comprises any one of SEQ ID NOS: 76-80, anyone of SEQ ID NOS: 88-90, and any one of SEQ ID NOS: 97-98. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 76-80, any one of SEQ ID NOS: 91-93, and any one ofSEQ ID NOS: 94-96. In some embodiments, an antibody or compound of thisdisclosure comprises any one of SEQ ID NOS: 76-80, any one of SEQ IDNOS: 91-93, and any one of SEQ ID NOS: 97-98. In some embodiments, anantibody or compound of this disclosure comprises any one of SEQ ID NOS:76-80, any one of SEQ ID NOS: 94-96, and any one of SEQ ID NOS: 97-98.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 81-87, any one of SEQ ID NOS: 88-90,and any one of SEQ ID NOS: 91-93. In some embodiments, an antibody orcompound of this disclosure comprises any one of SEQ ID NOS: 81-87, anyone of SEQ ID NOS: 88-90, and any one of SEQ ID NOS: 94-96. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 81-87, any one of SEQ ID NOS: 88-90, and any one ofSEQ ID NOS: 97-98. In some embodiments, an antibody or compound of thisdisclosure comprises any one of SEQ ID NOS: 81-87, any one of SEQ IDNOS: 91-93, and any one of SEQ ID NOS: 94-96. In some embodiments, anantibody or compound of this disclosure comprises any one of SEQ ID NOS:81-87, any one of SEQ ID NOS: 91-93, and any one of SEQ ID NOS: 97-98.In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 81-87, any one of SEQ ID NOS: 94-96,and any one of SEQ ID NOS: 97-98. In some embodiments, an antibody orcompound of this disclosure comprises any one of SEQ ID NOS: 88-90, anyone of SEQ ID NOS: 91-93, and any one of SEQ ID NOS: 94-96. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 88-90, any one of SEQ ID NOS: 91-93, and any one ofSEQ ID NOS: 97-98. In some embodiments, an antibody or compound of thisdisclosure comprises any one of SEQ ID NOS: 88-90, any one of SEQ IDNOS: 94-96, and any one of SEQ ID NOS: 97-98. In some embodiments, anantibody or compound of this disclosure comprises any one of SEQ ID NOS:91-93, any one of SEQ ID NOS: 94-96, and any one of SEQ ID NOS: 97-98.

In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 76-80, any one of SEQ ID NOS: 81-87,any one of SEQ ID NOS: 88-90, and any one of SEQ ID NOS: 91-93. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 76-80, any one of SEQ ID NOS: 81-87, any one of SEQID NOS: 88-90, and any one of SEQ ID NOS: 94-96. In some embodiments, anantibody or compound of this disclosure comprises any one of SEQ ID NOS:76-80, any one of SEQ ID NOS: 81-87, any one of SEQ ID NOS: 88-90, andany one of SEQ ID NOS: 97-98. In some embodiments, an antibody orcompound of this disclosure comprises any one of SEQ ID NOS: 76-80, anyone of SEQ ID NOS: 81-87, any one of SEQ ID NOS: 91-93, and any one ofSEQ ID NOS: 94-96. In some embodiments, an antibody or compound of thisdisclosure comprises any one of SEQ ID NOS: 76-80, any one of SEQ IDNOS: 81-87, any one of SEQ ID NOS: 91-93, and any one of SEQ ID NOS:97-98. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 76-80, any one of SEQ ID NOS: 81-87,any one of SEQ ID NOS: 94-96, and any one of SEQ ID NOS: 97-98. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 76-80, any one of SEQ ID NOS: 88-90, any one of SEQID NOS: 91-93, and any one of SEQ ID NOS: 94-96. In some embodiments, anantibody or compound of this disclosure comprises any one of SEQ ID NOS:76-80, any one of SEQ ID NOS: 88-90, any one of SEQ ID NOS: 91-93, andany one of SEQ ID NOS: 97-98. In some embodiments, an antibody orcompound of this disclosure comprises any one of SEQ ID NOS: 76-80, anyone of SEQ ID NOS: 88-90, any one of SEQ ID NOS: 94-96, and any one ofSEQ ID NOS: 97-98. In some embodiments, an antibody or compound of thisdisclosure comprises any one of SEQ ID NOS: 76-80, any one of SEQ IDNOS: 91-93, any one of SEQ ID NOS: 94-96, and any one of SEQ ID NOS:97-98. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 81-87, any one of SEQ ID NOS: 88-90,any one of SEQ ID NOS: 91-93, and any one of SEQ ID NOS: 94-96. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 81-87, any one of SEQ ID NOS: 88-90, any one of SEQID NOS: 91-93, and any one of SEQ ID NOS: 97-98. In some embodiments, anantibody or compound of this disclosure comprises any one of SEQ ID NOS:81-87, any one of SEQ ID NOS: 88-90, any one of SEQ ID NOS: 94-96, andany one of SEQ ID NOS: 97-98. In some embodiments, an antibody orcompound of this disclosure comprises any one of SEQ ID NOS: 81-87, anyone of SEQ ID NOS: 91-93, any one of SEQ ID NOS: 94-96, and any one ofSEQ ID NOS: 97-98. In some embodiments, an antibody or compound of thisdisclosure comprises any one of SEQ ID NOS: 88-90, any one of SEQ IDNOS: 91-93, any one of SEQ ID NOS: 94-96, and any one of SEQ ID NOS:97-98.

In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 76-80, any one of SEQ ID NOS: 81-87,any one of SEQ ID NOS: 88-90, any one of SEQ ID NOS: 91-93, and any oneof SEQ ID NOS: 94-96. In some embodiments, an antibody or compound ofthis disclosure comprises any one of SEQ ID NOS: 76-80, any one of SEQID NOS: 81-87, any one of SEQ ID NOS: 88-90, any one of SEQ ID NOS:91-93, and any one of SEQ ID NOS: 97-98. In some embodiments, anantibody or compound of this disclosure comprises any one of SEQ ID NOS:76-80, any one of SEQ ID NOS: 81-87, any one of SEQ ID NOS: 88-90, anyone of SEQ ID NOS: 94-96, and any one of SEQ ID NOS: 97-98. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 76-80, any one of SEQ ID NOS: 81-87, any one of SEQID NOS: 91-93, any one of SEQ ID NOS: 94-96, and any one of SEQ ID NOS:97-98. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 76-80, any one of SEQ ID NOS: 88-90,any one of SEQ ID NOS: 91-93, any one of SEQ ID NOS: 94-96, and any oneof SEQ ID NOS: 97-98. In some embodiments, an antibody or compound ofthis disclosure comprises any one of SEQ ID NOS: 81-87, any one of SEQID NOS: 88-90, any one of SEQ ID NOS: 91-93, any one of SEQ ID NOS:94-96, and any one of SEQ ID NOS: 97-98.

In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 76-80, any one of SEQ ID NOS: 81-87,any one of SEQ ID NOS: 88-90, any one of SEQ ID NOS: 91-93, any one ofSEQ ID NOS: 94-96, and any one of SEQ ID NOS: 97-98.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 76. In some embodiments, an antibody or compound ofthis disclosure comprises SEQ ID NO: 81. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 88. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 91. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 94. In some embodiments, an antibody or compound ofthis disclosure comprises SEQ ID NO: 97. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 77. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 82.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 76 and SEQ ID NO: 81. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 76 and SEQID NO: 88. In some embodiments, an antibody or compound of thisdisclosure comprises SEQ ID NO: 76 and SEQ ID NO: 91. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 76 and SEQ ID NO: 94. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 76 and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 81 and SEQ ID NO: 88. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 81 and SEQ ID NO: 91. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 81 and SEQ ID NO: 94. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 81 and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 88 and SEQ ID NO: 91. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 88 and SEQ ID NO: 94. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 88 and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 91 and SEQ ID NO: 94. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 91 and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 94 and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 77 and SEQ ID NO: 82. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 77 and SEQ ID NO: 88. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 77 and SEQ ID NO: 91. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 77 and SEQ ID NO: 94. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 77 and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 82 and SEQ ID NO: 88. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 82 and SEQ ID NO: 91. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 82 and SEQ ID NO: 94. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 82 and SEQ ID NO: 97.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 76, SEQ ID NO: 81, and SEQ ID NO: 88. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 76, SEQ ID NO: 81, and SEQ ID NO: 91. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 76, SEQ IDNO: 81, and SEQ ID NO: 94. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 76, SEQ ID NO: 81, and SEQ IDNO: 97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 76, SEQ ID NO: 88, and SEQ ID NO: 91. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 76, SEQ ID NO: 88, and SEQ ID NO: 94. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 76, SEQ IDNO: 88, and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 76, SEQ ID NO: 91, and SEQ IDNO: 94. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 76, SEQ ID NO: 91, and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 76, SEQ ID NO: 94, and SEQ ID NO: 97. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 81, SEQ IDNO: 88, and SEQ ID NO: 91. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 81, SEQ ID NO: 88, and SEQ IDNO: 94. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 81, SEQ ID NO: 88, and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 81, SEQ ID NO: 91, and SEQ ID NO: 94. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 81, SEQ IDNO: 91, and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 81, SEQ ID NO: 94, and SEQ IDNO: 97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 88, SEQ ID NO: 91, and SEQ ID NO: 94. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 88, SEQ ID NO: 91, and SEQ ID NO: 97. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 88, SEQ IDNO: 94, and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 91, SEQ ID NO: 94, and SEQ IDNO: 97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 77, SEQ ID NO: 82, and SEQ ID NO: 88. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 77, SEQ ID NO: 82, and SEQ ID NO: 91. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 77, SEQ IDNO: 82, and SEQ ID NO: 94. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 77, SEQ ID NO: 82, and SEQ IDNO: 97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 77, SEQ ID NO: 88, and SEQ ID NO: 91. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 77, SEQ ID NO: 88, and SEQ ID NO: 94. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 77, SEQ IDNO: 88, and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 77, SEQ ID NO: 91, and SEQ IDNO: 94. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 77, SEQ ID NO: 91, and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 77, SEQ ID NO: 94, and SEQ ID NO: 97. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 82, SEQ IDNO: 88, and SEQ ID NO: 91. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 82, SEQ ID NO: 88, and SEQ IDNO: 94. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 82, SEQ ID NO: 88, and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 82, SEQ ID NO: 91, and SEQ ID NO: 94. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 82, SEQ IDNO: 91, and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 82, SEQ ID NO: 94, and SEQ IDNO: 97.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 76, SEQ ID NO: 81, SEQ ID NO: 88, and SEQ ID NO:91. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 76, SEQ ID NO: 81, SEQ ID NO: 88, and SEQ ID NO:94. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 76, SEQ ID NO: 81, SEQ ID NO: 88, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 76, SEQ ID NO: 81, SEQ ID NO: 91, and SEQ ID NO:94. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 76, SEQ ID NO: 81, SEQ ID NO: 91, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 76, SEQ ID NO: 81, SEQ ID NO: 94, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 76, SEQ ID NO: 88, SEQ ID NO: 91, and SEQ ID NO:94. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 76, SEQ ID NO: 88, SEQ ID NO: 91, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 76, SEQ ID NO: 88, SEQ ID NO: 94, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 76, SEQ ID NO: 91, SEQ ID NO: 94, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 91, and SEQ ID NO:94. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 91, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 94, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 81, SEQ ID NO: 91, SEQ ID NO: 94, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 88, SEQ ID NO: 91, SEQ ID NO: 94, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 77, SEQ ID NO: 82, SEQ ID NO: 88, and SEQ ID NO:91. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 77, SEQ ID NO: 82, SEQ ID NO: 88, and SEQ ID NO:94. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 77, SEQ ID NO: 82, SEQ ID NO: 88, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 77, SEQ ID NO: 82, SEQ ID NO: 91, and SEQ ID NO:94. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 77, SEQ ID NO: 82, SEQ ID NO: 91, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 77, SEQ ID NO: 82, SEQ ID NO: 94, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 77, SEQ ID NO: 88, SEQ ID NO: 91, and SEQ ID NO:94. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 77, SEQ ID NO: 88, SEQ ID NO: 91, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 77, SEQ ID NO: 88, SEQ ID NO: 94, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 77, SEQ ID NO: 91, SEQ ID NO: 94, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 82, SEQ ID NO: 88, SEQ ID NO: 91, and SEQ ID NO:94. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 82, SEQ ID NO: 88, SEQ ID NO: 91, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 82, SEQ ID NO: 88, SEQ ID NO: 94, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 82, SEQ ID NO: 91, SEQ ID NO: 94, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 88, SEQ ID NO: 91, SEQ ID NO: 94, and SEQ ID NO:97.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 76, SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 91,and SEQ ID NO: 94. In some embodiments, an antibody or compound of thisdisclosure comprises SEQ ID NO: 76, SEQ ID NO: 81, SEQ ID NO: 88, SEQ IDNO: 91, and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 76, SEQ ID NO: 81, SEQ ID NO:88, SEQ ID NO: 94, and SEQ ID NO: 97. In some embodiments, an antibodyor compound of this disclosure comprises SEQ ID NO: 76, SEQ ID NO: 81,SEQ ID NO: 91, SEQ ID NO: 94, and SEQ ID NO: 97. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 76, SEQ IDNO: 88, SEQ ID NO: 91, SEQ ID NO: 94, and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 81, SEQ ID NO: 88, SEQ ID NO: 91, SEQ ID NO: 94, and SEQ ID NO: 97.In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 77, SEQ ID NO: 82, SEQ ID NO: 88, SEQ ID NO: 91,and SEQ ID NO: 94. In some embodiments, an antibody or compound of thisdisclosure comprises SEQ ID NO: 77, SEQ ID NO: 82, SEQ ID NO: 88, SEQ IDNO: 91, and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 77, SEQ ID NO: 82, SEQ ID NO:88, SEQ ID NO: 94, and SEQ ID NO: 97. In some embodiments, an antibodyor compound of this disclosure comprises SEQ ID NO: 77, SEQ ID NO: 82,SEQ ID NO: 91, SEQ ID NO: 94, and SEQ ID NO: 97. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 82, SEQ IDNO: 88, SEQ ID NO: 91, SEQ ID NO: 94, and SEQ ID NO: 97.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 76, SEQ ID NO: 81, SEQ ID NO: 88, SEQ ID NO: 91,SEQ ID NO: 94, and SEQ ID NO: 97. In some embodiments, an antibody orcompound of this disclosure comprises SEQ ID NO: 77, SEQ ID NO: 82, SEQID NO: 88, SEQ ID NO: 91, SEQ ID NO: 94, and SEQ ID NO: 97.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 78. In some embodiments, an antibody or compound ofthis disclosure comprises SEQ ID NO: 83. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 89. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 92. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 95. In some embodiments, an antibody or compound ofthis disclosure comprises SEQ ID NO: 97.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 78 and SEQ ID NO: 83. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 78 and SEQID NO: 89. In some embodiments, an antibody or compound of thisdisclosure comprises SEQ ID NO: 78 and SEQ ID NO: 92. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 78 and SEQ ID NO: 95. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 78 and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 83 and SEQ ID NO: 89. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 83 and SEQ ID NO: 92. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 83 and SEQ ID NO: 95. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 83 and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 89 and SEQ ID NO: 92. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 89 and SEQ ID NO: 95. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 89 and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 92 and SEQ ID NO: 95. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 92 and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 95 and SEQ ID NO: 97.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 78, SEQ ID NO: 83, and SEQ ID NO: 89. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 78, SEQ ID NO: 83, and SEQ ID NO: 92. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 78, SEQ IDNO: 83, and SEQ ID NO: 95. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 78, SEQ ID NO: 83, and SEQ IDNO: 97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 78, SEQ ID NO: 89, and SEQ ID NO: 92. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 78, SEQ ID NO: 89, and SEQ ID NO: 95. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 78, SEQ IDNO: 89, and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 78, SEQ ID NO: 92, and SEQ IDNO: 95. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 78, SEQ ID NO: 92, and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 78, SEQ ID NO: 95, and SEQ ID NO: 97. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 83, SEQ IDNO: 89, and SEQ ID NO: 92. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 83, SEQ ID NO: 89, and SEQ IDNO: 95. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 83, SEQ ID NO: 89, and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 83, SEQ ID NO: 92, and SEQ ID NO: 95. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 83, SEQ IDNO: 92, and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 83, SEQ ID NO: 95, and SEQ IDNO: 97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 89, SEQ ID NO: 92, and SEQ ID NO: 95. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 89, SEQ ID NO: 92, and SEQ ID NO: 97. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 89, SEQ IDNO: 95, and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 92, SEQ ID NO: 95, and SEQ IDNO: 97.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 78, SEQ ID NO: 83, SEQ ID NO: 89, and SEQ ID NO:92. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 78, SEQ ID NO: 83, SEQ ID NO: 89, and SEQ ID NO:95. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 78, SEQ ID NO: 83, SEQ ID NO: 89, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 78, SEQ ID NO: 83, SEQ ID NO: 92, and SEQ ID NO:95. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 78, SEQ ID NO: 83, SEQ ID NO: 92, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 78, SEQ ID NO: 83, SEQ ID NO: 95, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 78, SEQ ID NO: 89, SEQ ID NO: 92, and SEQ ID NO:95. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 78, SEQ ID NO: 89, SEQ ID NO: 92, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 78, SEQ ID NO: 89, SEQ ID NO: 95, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 78, SEQ ID NO: 92, SEQ ID NO: 95, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 83, SEQ ID NO: 89, SEQ ID NO: 92, and SEQ ID NO:95. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 83, SEQ ID NO: 89, SEQ ID NO: 92, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 83, SEQ ID NO: 89, SEQ ID NO: 95, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 83, SEQ ID NO: 92, SEQ ID NO: 95, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 89, SEQ ID NO: 92, SEQ ID NO: 95, and SEQ ID NO:97.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 78, SEQ ID NO: 83, SEQ ID NO: 89, SEQ ID NO: 92,and SEQ ID NO: 95. In some embodiments, an antibody or compound of thisdisclosure comprises SEQ ID NO: 78, SEQ ID NO: 83, SEQ ID NO: 89, SEQ IDNO: 92, and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 78, SEQ ID NO: 83, SEQ ID NO:89, SEQ ID NO: 95, and SEQ ID NO: 97. In some embodiments, an antibodyor compound of this disclosure comprises SEQ ID NO: 78, SEQ ID NO: 83,SEQ ID NO: 92, SEQ ID NO: 95, and SEQ ID NO: 97. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 78, SEQ IDNO: 89, SEQ ID NO: 92, SEQ ID NO: 95, and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 83, SEQ ID NO: 89, SEQ ID NO: 92, SEQ ID NO: 95, and SEQ ID NO: 97.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 78, SEQ ID NO: 83, SEQ ID NO: 89, SEQ ID NO: 92,SEQ ID NO: 95, and SEQ ID NO: 97.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79. In some embodiments, an antibody or compound ofthis disclosure comprises SEQ ID NO: 84. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 90. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 93. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 96. In some embodiments, an antibody or compound ofthis disclosure comprises SEQ ID NO: 98. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 86.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79 and SEQ ID NO: 84. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 79 and SEQID NO: 90. In some embodiments, an antibody or compound of thisdisclosure comprises SEQ ID NO: 79 and SEQ ID NO: 93. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 79 and SEQ ID NO: 96. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 79 and SEQ ID NO: 98. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 84 and SEQ ID NO: 90. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 84 and SEQ ID NO: 93. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 84 and SEQ ID NO: 96. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 84 and SEQ ID NO: 98. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 90 and SEQ ID NO: 93. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 90 and SEQ ID NO: 96. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 90 and SEQ ID NO: 98. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 93 and SEQ ID NO: 96. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 93 and SEQ ID NO: 98. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 96 and SEQ ID NO: 98. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 79 and SEQ ID NO: 86. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 86 and SEQ ID NO: 90. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 86 and SEQ ID NO: 93. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 86 and SEQ ID NO: 96. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 86 and SEQ ID NO: 98.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 84, and SEQ ID NO: 90. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 79, SEQ ID NO: 84, and SEQ ID NO: 93. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 79, SEQ IDNO: 84, and SEQ ID NO: 96. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 79, SEQ ID NO: 84, and SEQ IDNO: 98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 90, and SEQ ID NO: 93. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 79, SEQ ID NO: 90, and SEQ ID NO: 96. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 79, SEQ IDNO: 90, and SEQ ID NO: 98. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 79, SEQ ID NO: 93, and SEQ IDNO: 96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 93, and SEQ ID NO: 98. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 79, SEQ ID NO: 96, and SEQ ID NO: 98. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 84, SEQ IDNO: 90, and SEQ ID NO: 93. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 84, SEQ ID NO: 90, and SEQ IDNO: 96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 84, SEQ ID NO: 90, and SEQ ID NO: 98. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 84, SEQ ID NO: 93, and SEQ ID NO: 96. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 84, SEQ IDNO: 93, and SEQ ID NO: 98. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 84, SEQ ID NO: 96, and SEQ IDNO: 98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 90, SEQ ID NO: 93, and SEQ ID NO: 96. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 90, SEQ ID NO: 93, and SEQ ID NO: 98. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 90, SEQ IDNO: 96, and SEQ ID NO: 98. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 93, SEQ ID NO: 96, and SEQ IDNO: 98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 86, and SEQ ID NO: 90. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 79, SEQ ID NO: 86, and SEQ ID NO: 93. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 79, SEQ IDNO: 86, and SEQ ID NO: 96. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 79, SEQ ID NO: 86, and SEQ IDNO: 98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 86, SEQ ID NO: 90, and SEQ ID NO: 93. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 86, SEQ ID NO: 90, and SEQ ID NO: 96. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 86, SEQ IDNO: 90, and SEQ ID NO: 98. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 86, SEQ ID NO: 93, and SEQ IDNO: 96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 86, SEQ ID NO: 93, and SEQ ID NO: 98. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 86, SEQ ID NO: 96, and SEQ ID NO: 98.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 84, SEQ ID NO: 90, and SEQ ID NO:93. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 84, SEQ ID NO: 90, and SEQ ID NO:96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 84, SEQ ID NO: 90, and SEQ ID NO:98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 84, SEQ ID NO: 93, and SEQ ID NO:96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 84, SEQ ID NO: 93, and SEQ ID NO:98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 84, SEQ ID NO: 96, and SEQ ID NO:98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 90, SEQ ID NO: 93, and SEQ ID NO:96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 90, SEQ ID NO: 93, and SEQ ID NO:98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 90, SEQ ID NO: 96, and SEQ ID NO:98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 93, SEQ ID NO: 96, and SEQ ID NO:98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 84, SEQ ID NO: 90, SEQ ID NO: 93, and SEQ ID NO:96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 84, SEQ ID NO: 90, SEQ ID NO: 93, and SEQ ID NO:98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 84, SEQ ID NO: 90, SEQ ID NO: 96, and SEQ ID NO:98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 84, SEQ ID NO: 93, SEQ ID NO: 96, and SEQ ID NO:98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 90, SEQ ID NO: 93, SEQ ID NO: 96, and SEQ ID NO:98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 86, SEQ ID NO: 90, and SEQ ID NO:93. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 86, SEQ ID NO: 90, and SEQ ID NO:96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 86, SEQ ID NO: 90, and SEQ ID NO:98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 86, SEQ ID NO: 93, and SEQ ID NO:96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 86, SEQ ID NO: 93, and SEQ ID NO:98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 86, SEQ ID NO: 96, and SEQ ID NO:98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 86, SEQ ID NO: 90, SEQ ID NO: 93, and SEQ ID NO:96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 86, SEQ ID NO: 90, SEQ ID NO: 93, and SEQ ID NO:98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 86, SEQ ID NO: 90, SEQ ID NO: 96, and SEQ ID NO:98. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 86, SEQ ID NO: 93, SEQ ID NO: 96, and SEQ ID NO:98.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 84, SEQ ID NO: 90, SEQ ID NO: 93,and SEQ ID NO: 96. In some embodiments, an antibody or compound of thisdisclosure comprises SEQ ID NO: 79, SEQ ID NO: 84, SEQ ID NO: 90, SEQ IDNO: 93, and SEQ ID NO: 98. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 79, SEQ ID NO: 84, SEQ ID NO:90, SEQ ID NO: 96, and SEQ ID NO: 98. In some embodiments, an antibodyor compound of this disclosure comprises SEQ ID NO: 79, SEQ ID NO: 84,SEQ ID NO: 93, SEQ ID NO: 96, and SEQ ID NO: 98. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 79, SEQ IDNO: 90, SEQ ID NO: 93, SEQ ID NO: 96, and SEQ ID NO: 98. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 84, SEQ ID NO: 90, SEQ ID NO: 93, SEQ ID NO: 96, and SEQ ID NO: 98.In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 86, SEQ ID NO: 90, SEQ ID NO: 93,and SEQ ID NO: 96. In some embodiments, an antibody or compound of thisdisclosure comprises SEQ ID NO: 79, SEQ ID NO: 86, SEQ ID NO: 90, SEQ IDNO: 93, and SEQ ID NO: 98. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 79, SEQ ID NO: 86, SEQ ID NO:90, SEQ ID NO: 96, and SEQ ID NO: 98. In some embodiments, an antibodyor compound of this disclosure comprises SEQ ID NO: 79, SEQ ID NO: 86,SEQ ID NO: 93, SEQ ID NO: 96, and SEQ ID NO: 98. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 86, SEQ IDNO: 90, SEQ ID NO: 93, SEQ ID NO: 96, and SEQ ID NO: 98.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 79, SEQ ID NO: 84, SEQ ID NO: 90, SEQ ID NO: 93,SEQ ID NO: 96, and SEQ ID NO: 98. In some embodiments, an antibody orcompound of this disclosure comprises SEQ ID NO: 79, SEQ ID NO: 86, SEQID NO: 90, SEQ ID NO: 93, SEQ ID NO: 96, and SEQ ID NO: 98.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80. In some embodiments, an antibody or compound ofthis disclosure comprises SEQ ID NO: 85. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 89. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 91. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 96. In some embodiments, an antibody or compound ofthis disclosure comprises SEQ ID NO: 97. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 87.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80 and SEQ ID NO: 85. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 80 and SEQID NO: 89. In some embodiments, an antibody or compound of thisdisclosure comprises SEQ ID NO: 80 and SEQ ID NO: 91. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 80 and SEQ ID NO: 96. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 80 and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 85 and SEQ ID NO: 89. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 85 and SEQ ID NO: 91. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 85 and SEQ ID NO: 96. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 85 and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 89 and SEQ ID NO: 91. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 89 and SEQ ID NO: 96. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 89 and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 91 and SEQ ID NO: 96. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 91 and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 96 and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 80 and SEQ ID NO: 87. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 87 and SEQ ID NO: 89. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 87 and SEQ ID NO: 91. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 87 and SEQ ID NO: 96. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 87 and SEQ ID NO: 97.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 85, and SEQ ID NO: 89. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 80, SEQ ID NO: 85, and SEQ ID NO: 91. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 80, SEQ IDNO: 85, and SEQ ID NO: 96. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 80, SEQ ID NO: 85, and SEQ IDNO: 97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 89, and SEQ ID NO: 91. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 80, SEQ ID NO: 89, and SEQ ID NO: 96. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 80, SEQ IDNO: 89, and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 80, SEQ ID NO: 91, and SEQ IDNO: 96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 91, and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 80, SEQ ID NO: 96, and SEQ ID NO: 97. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 85, SEQ IDNO: 89, and SEQ ID NO: 91. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 85, SEQ ID NO: 89, and SEQ IDNO: 96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 85, SEQ ID NO: 89, and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 85, SEQ ID NO: 91, and SEQ ID NO: 96. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 85, SEQ IDNO: 91, and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 85, SEQ ID NO: 96, and SEQ IDNO: 97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 89, SEQ ID NO: 91, and SEQ ID NO: 96. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 89, SEQ ID NO: 91, and SEQ ID NO: 97. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 89, SEQ IDNO: 96, and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 91, SEQ ID NO: 96, and SEQ IDNO: 97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 87, and SEQ ID NO: 89. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 80, SEQ ID NO: 87, and SEQ ID NO: 91. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 80, SEQ IDNO: 87, and SEQ ID NO: 96. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 80, SEQ ID NO: 87, and SEQ IDNO: 97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 87, SEQ ID NO: 89, and SEQ ID NO: 91. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 87, SEQ ID NO: 89, and SEQ ID NO: 96. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 87, SEQ IDNO: 89, and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 87, SEQ ID NO: 91, and SEQ IDNO: 96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 87, SEQ ID NO: 91, and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 87, SEQ ID NO: 96, and SEQ ID NO: 97.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 85, SEQ ID NO: 89, and SEQ ID NO:91. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 85, SEQ ID NO: 89, and SEQ ID NO:96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 85, SEQ ID NO: 89, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 85, SEQ ID NO: 91, and SEQ ID NO:96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 85, SEQ ID NO: 91, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 85, SEQ ID NO: 96, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 89, SEQ ID NO: 91, and SEQ ID NO:96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 89, SEQ ID NO: 91, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 89, SEQ ID NO: 96, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 91, SEQ ID NO: 96, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 85, SEQ ID NO: 89, SEQ ID NO: 91, and SEQ ID NO:96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 85, SEQ ID NO: 89, SEQ ID NO: 91, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 85, SEQ ID NO: 89, SEQ ID NO: 96, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 85, SEQ ID NO: 91, SEQ ID NO: 96, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 96, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 87, SEQ ID NO: 89, and SEQ ID NO:91. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 87, SEQ ID NO: 89, and SEQ ID NO:96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 87, SEQ ID NO: 89, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 87, SEQ ID NO: 91, and SEQ ID NO:96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 87, SEQ ID NO: 91, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 87, SEQ ID NO: 96, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, and SEQ ID NO:96. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 96, and SEQ ID NO:97. In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 87, SEQ ID NO: 91, SEQ ID NO: 96, and SEQ ID NO:97.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 85, SEQ ID NO: 89, SEQ ID NO: 91,and SEQ ID NO: 96. In some embodiments, an antibody or compound of thisdisclosure comprises SEQ ID NO: 80, SEQ ID NO: 85, SEQ ID NO: 89, SEQ IDNO: 91, and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 80, SEQ ID NO: 85, SEQ ID NO:89, SEQ ID NO: 96, and SEQ ID NO: 97. In some embodiments, an antibodyor compound of this disclosure comprises SEQ ID NO: 80, SEQ ID NO: 85,SEQ ID NO: 91, SEQ ID NO: 96, and SEQ ID NO: 97. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 80, SEQ IDNO: 89, SEQ ID NO: 91, SEQ ID NO: 96, and SEQ ID NO: 97. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 85, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 96, and SEQ ID NO: 97.In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91,and SEQ ID NO: 96. In some embodiments, an antibody or compound of thisdisclosure comprises SEQ ID NO: 80, SEQ ID NO: 87, SEQ ID NO: 89, SEQ IDNO: 91, and SEQ ID NO: 97. In some embodiments, an antibody or compoundof this disclosure comprises SEQ ID NO: 80, SEQ ID NO: 87, SEQ ID NO:89, SEQ ID NO: 96, and SEQ ID NO: 97. In some embodiments, an antibodyor compound of this disclosure comprises SEQ ID NO: 80, SEQ ID NO: 87,SEQ ID NO: 91, SEQ ID NO: 96, and SEQ ID NO: 97. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 87, SEQ IDNO: 89, SEQ ID NO: 91, SEQ ID NO: 96, and SEQ ID NO: 97.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 85, SEQ ID NO: 89, SEQ ID NO: 91,SEQ ID NO: 96, and SEQ ID NO: 97. In some embodiments, an antibody orcompound of this disclosure comprises SEQ ID NO: 80, SEQ ID NO: 87, SEQID NO: 89, SEQ ID NO: 91, SEQ ID NO: 96, and SEQ ID NO: 97.

In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 99-113. In some embodiments, anantibody or compound of this disclosure comprises any one of SEQ ID NOS:114-123. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 124-131. In some embodiments, anantibody or compound of this disclosure comprises any one of SEQ ID NOS:132-137. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 138-143. In some embodiments, anantibody or compound of this disclosure comprises any one of SEQ ID NOS:144-146.

In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 99-113 and any one of SEQ ID NOS:114-123. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 99-113 and any one of SEQ ID NOS:124-131. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 99-113 and any one of SEQ ID NOS:132-137. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 99-113 and any one of SEQ ID NOS:138-143. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 99-113 and any one of SEQ ID NOS:144-146. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 114-123 and any one of SEQ ID NOS:124-131. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 114-123 and any one of SEQ ID NOS:132-137. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 114-123 and any one of SEQ ID NOS:138-143. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 114-123 and any one of SEQ ID NOS:144-146. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 124-131 and any one of SEQ ID NOS:132-137. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 124-131 and any one of SEQ ID NOS:138-143. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 124-131 and any one of SEQ ID NOS:144-146. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 132-137 and any one of SEQ ID NOS:138-143. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 132-137 and any one of SEQ ID NOS:144-146. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 138-143 and any one of SEQ ID NOS:144-146.

In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 114-123,and any one of SEQ ID NOS: 124-131. In some embodiments, an antibody orcompound of this disclosure comprises any one of SEQ ID NOS: 99-113, anyone of SEQ ID NOS: 114-123, and any one of SEQ ID NOS: 132-137. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 114-123, and any oneof SEQ ID NOS: 138-143. In some embodiments, an antibody or compound ofthis disclosure comprises any one of SEQ ID NOS: 99-113, any one of SEQID NOS: 114-123, and any one of SEQ ID NOS: 144-146. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 124-131, and any oneof SEQ ID NOS: 132-137. In some embodiments, an antibody or compound ofthis disclosure comprises any one of SEQ ID NOS: 99-113, any one of SEQID NOS: 124-131, and any one of SEQ ID NOS: 138-143. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 124-131, and any oneof SEQ ID NOS: 144-146. In some embodiments, an antibody or compound ofthis disclosure comprises any one of SEQ ID NOS: 99-113, any one of SEQID NOS: 132-137, and any one of SEQ ID NOS: 138-143. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 132-137, and any oneof SEQ ID NOS: 144-146. In some embodiments, an antibody or compound ofthis disclosure comprises any one of SEQ ID NOS: 99-113, any one of SEQID NOS: 138-143, and any one of SEQ ID NOS: 144-146. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 114-123, any one of SEQ ID NOS: 124-131, and any oneof SEQ ID NOS: 132-137. In some embodiments, an antibody or compound ofthis disclosure comprises any one of SEQ ID NOS: 114-123, any one of SEQID NOS: 124-131, and any one of SEQ ID NOS: 138-143. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 114-123, any one of SEQ ID NOS: 124-131, and any oneof SEQ ID NOS: 144-146. In some embodiments, an antibody or compound ofthis disclosure comprises any one of SEQ ID NOS: 114-123, any one of SEQID NOS: 132-137, and any one of SEQ ID NOS: 138-143. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 114-123, any one of SEQ ID NOS: 132-137, and any oneof SEQ ID NOS: 144-146. In some embodiments, an antibody or compound ofthis disclosure comprises any one of SEQ ID NOS: 114-123, any one of SEQID NOS: 138-143, and any one of SEQ ID NOS: 144-146. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 124-131, any one of SEQ ID NOS: 132-137, and any oneof SEQ ID NOS: 138-143. In some embodiments, an antibody or compound ofthis disclosure comprises any one of SEQ ID NOS: 124-131, any one of SEQID NOS: 132-137, and any one of SEQ ID NOS: 144-146. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 124-131, any one of SEQ ID NOS: 138-143, and any oneof SEQ ID NOS: 144-146. In some embodiments, an antibody or compound ofthis disclosure comprises any one of SEQ ID NOS: 132-137, any one of SEQID NOS: 138-143, and any one of SEQ ID NOS: 144-146.

In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 114-123,any one of SEQ ID NOS: 124-131, and any one of SEQ ID NOS: 132-137. Insome embodiments, an antibody or compound of this disclosure comprisesany one of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 114-123, any oneof SEQ ID NOS: 124-131, and any one of SEQ ID NOS: 138-143. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 114-123, any one ofSEQ ID NOS: 124-131, and any one of SEQ ID NOS: 144-146. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 114-123, any one ofSEQ ID NOS: 132-137, and any one of SEQ ID NOS: 138-143. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 114-123, any one ofSEQ ID NOS: 132-137, and any one of SEQ ID NOS: 144-146. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 114-123, any one ofSEQ ID NOS: 138-143, and any one of SEQ ID NOS: 144-146. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 124-131, any one ofSEQ ID NOS: 132-137, and any one of SEQ ID NOS: 138-143. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 124-131, any one ofSEQ ID NOS: 132-137, and any one of SEQ ID NOS: 144-146. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 124-131, any one ofSEQ ID NOS: 138-143, and any one of SEQ ID NOS: 144-146. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 132-137, any one ofSEQ ID NOS: 138-143, and any one of SEQ ID NOS: 144-146. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 114-123, any one of SEQ ID NOS: 124-131, any one ofSEQ ID NOS: 132-137, and any one of SEQ ID NOS: 138-143. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 114-123, any one of SEQ ID NOS: 124-131, any one ofSEQ ID NOS: 132-137, and any one of SEQ ID NOS: 144-146. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 114-123, any one of SEQ ID NOS: 124-131, any one ofSEQ ID NOS: 138-143, and any one of SEQ ID NOS: 144-146. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 114-123, any one of SEQ ID NOS: 132-137, any one ofSEQ ID NOS: 138-143, and any one of SEQ ID NOS: 144-146. In someembodiments, an antibody or compound of this disclosure comprises anyone of SEQ ID NOS: 124-131, any one of SEQ ID NOS: 132-137, any one ofSEQ ID NOS: 138-143, and any one of SEQ ID NOS: 144-146.

In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 114-123,any one of SEQ ID NOS: 124-131, any one of SEQ ID NOS: 132-137, and anyone of SEQ ID NOS: 138-143. In some embodiments, an antibody or compoundof this disclosure comprises any one of SEQ ID NOS: 99-113, any one ofSEQ ID NOS: 114-123, any one of SEQ ID NOS: 124-131, any one of SEQ IDNOS: 132-137, and any one of SEQ ID NOS: 144-146. In some embodiments,an antibody or compound of this disclosure comprises any one of SEQ IDNOS: 99-113, any one of SEQ ID NOS: 114-123, any one of SEQ ID NOS:124-131, any one of SEQ ID NOS: 138-143, and any one of SEQ ID NOS:144-146. In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 114-123,any one of SEQ ID NOS: 132-137, any one of SEQ ID NOS: 138-143, and anyone of SEQ ID NOS: 144-146. In some embodiments, an antibody or compoundof this disclosure comprises any one of SEQ ID NOS: 99-113, any one ofSEQ ID NOS: 124-131, any one of SEQ ID NOS: 132-137, any one of SEQ IDNOS: 138-143, and any one of SEQ ID NOS: 144-146. In some embodiments,an antibody or compound of this disclosure comprises any one of SEQ IDNOS: 114-123, any one of SEQ ID NOS: 124-131, any one of SEQ ID NOS:132-137, any one of SEQ ID NOS: 138-143, and any one of SEQ ID NOS:144-146.

In some embodiments, an antibody or compound of this disclosurecomprises any one of SEQ ID NOS: 99-113, any one of SEQ ID NOS: 114-123,any one of SEQ ID NOS: 124-131, any one of SEQ ID NOS: 132-137, any oneof SEQ ID NOS: 138-143, and any one of SEQ ID NOS: 144-146.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO 103, SEQ ID NO: 118, SEQ ID NO: 125, SEQ ID NO: 132,SEQ ID NO: 140, and SEQ ID NO: 144. In some embodiments, an antibody orcompound of this disclosure comprises SEQ ID NO 108, SEQ ID NO: 123, SEQID NO: 127, SEQ ID NO: 135, SEQ ID NO: 143, and SEQ ID NO: 145. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO 113, SEQ ID NO: 123, SEQ ID NO: 130, SEQ ID NO: 135, SEQ ID NO: 143,and SEQ ID NO: 145.

In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 85, SEQ ID NO: 89, SEQ ID NO: 91,SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO 103, SEQ ID NO: 118, SEQ ID NO:125, SEQ ID NO: 132, SEQ ID NO: 140, and SEQ ID NO: 144. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 80, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 96, SEQID NO: 97, SEQ ID NO 103, SEQ ID NO: 118, SEQ ID NO: 125, SEQ ID NO:132, SEQ ID NO: 140, and SEQ ID NO: 144. In some embodiments, anantibody or compound of this disclosure comprises SEQ ID NO: 80, SEQ IDNO: 85, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 96, SEQ ID NO: 97, SEQID NO 108, SEQ ID NO: 123, SEQ ID NO: 127, SEQ ID NO: 135, SEQ ID NO:143, and SEQ ID NO: 145. In some embodiments, an antibody or compound ofthis disclosure comprises SEQ ID NO: 80, SEQ ID NO: 87, SEQ ID NO: 89,SEQ ID NO: 91, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO 108, SEQ ID NO:123, SEQ ID NO: 127, SEQ ID NO: 135, SEQ ID NO: 143, and SEQ ID NO: 145.In some embodiments, an antibody or compound of this disclosurecomprises SEQ ID NO: 80, SEQ ID NO: 85, SEQ ID NO: 89, SEQ ID NO: 91,SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO 113, SEQ ID NO: 123, SEQ ID NO:130, SEQ ID NO: 135, SEQ ID NO: 143, and SEQ ID NO: 145. In someembodiments, an antibody or compound of this disclosure comprises SEQ IDNO: 80, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 96, SEQID NO: 97, SEQ ID NO 113, SEQ ID NO: 123, SEQ ID NO: 130, SEQ ID NO:135, SEQ ID NO: 143, and SEQ ID NO: 145.

EXAMPLES Example 1: A Tetravalent Bispecific Antibody ComprisingAntibody HC2:LC and Aflibercept-Derived Sequences

To generate a tetravalent bispecific antibody in which the heavy chainof antibody HC2:LC1 was fused to an aflibercept-derived VEGF-bindingdomain, an amino acid sequence was generated comprising the followingappended amino acid sequences, from N-terminus to C-terminus:

1) SEQ ID NO: 14 (heavy chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 22 (aflibercept-derived sequence).

The resulting polypeptide (SEQ ID NO: 149) was co-expressed with SEQ IDNO: 17, to provide tetravalent, bispecific antibody HC2-AFL:LC1,comprising the sequences shown in TABLE 33. Amino acids 1-19 of SEQ IDNO: 149 are the heavy chain signal peptide (SEQ ID NO: 11). Amino acids1-20 of SEQ ID NO: 17 are the light chain signal peptide (SEQ ID NO:12). In some embodiments, a mature HC2-AFL:LC1 does not comprise thesignal peptides. For example, a mature HC2-AFL:LC1 of the disclosure cancomprise SEQ ID NO: 254 and SEQ ID NO: 250.

TABLE 33 SEQ ID NO: Name Amino acid sequence 149 signalMGWTLVFLFLLSVTAGVHSEVQLVESGGGLVQPGGSLRLSC peptide-AASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATY HC2-YAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRD AFLYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKGGGGSSDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAAS SGLMTKKNSTFVRVHEK 17 SignalMVSSAQFLGLLLLCFQGTRCDVVMTQSPSFLSASVGDRVTIT peptide-CKASQHVGTAVAWYQQRPGKAPKLLIYWASTRHTGVPSRF LC1SGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGEC 254HC2- EVQLVESGGGLVQPGGSLRLSCAASGFTFNANAMNWVRQA AFLPGKGLEWVGRIRTKSNNYATYYAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRDYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKGGGGSSDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEK

Example 2: A Tetravalent Bispecific Antibody Comprising Antibody HC2:LC1and Brolucizumab-Derived Sequences

To generate a tetravalent bispecific antibody in which the heavy chainof antibody HC2:LC1 was fused to a brolucizumab-derived VEGF-bindingdomain, an amino acid sequence was generated comprising the followingappended amino acid sequences, from N-terminus to C-terminus:

1) SEQ ID NO: 14 (heavy chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 23 (brolucizumab-derived sequence).

The resulting polypeptide (SEQ ID NO: 150) was co-expressed with SEQ IDNO: 17, to provide tetravalent, bispecific antibody HC2-BRO:LC1,comprising the sequences shown in TABLE 34. Amino acids 1-19 of SEQ IDNO: 150 are the heavy chain signal peptide (SEQ ID NO: 11). Amino acids1-20 of SEQ ID NO: 17 are the light chain signal peptide (SEQ ID NO:12). In some embodiments, a mature HC2-BRO:LC1 does not comprise thesignal peptides. For example, a mature HC2-BRO:LC1 of the disclosure cancomprise SEQ ID NO: 255 and SEQ ID NO: 250.

TABLE 34 SEQ ID NO: Name Amino acid sequence 150 signalMGWTLVFLFLLSVTAGVHSEVQLVESGGGLVQPGGSLRLSC peptide-AASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATY HC2-YAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRD BROYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKGGGGSEIVMTQSPSTLSASVGDRVIITCQASEIIHSWLAWYQQKPGKAPKLLIYLASTLASGVPSRFSGSGSGAEFTLTISSLQPDDFATYYCQNVYLASTNGANFGQGTKLTVLGGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCTASGFSLTDYYYMTWVRQAPGKGLEWVGFIDPDDDPYYATWAKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCAGGDHNSGWGLDIWGQGTLVTVSS 17Signal MVSSAQFLGLLLLCFQGTRCDVVMTQSPSFLSASVGDRVTIT peptide-CKASQHVGTAVAWYQQRPGKAPKLLIYWASTRHTGVPSRF LC1SGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGEC 255HC2- EVQLVESGGGLVQPGGSLRLSCAASGFTFNANAMNWVRQA BROPGKGLEWVGRIRTKSNNYATYYAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRDYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKGGGGSEIVMTQSPSTLSASVGDRVIITCQASEIIHSWLAWYQQKPGKAPKLLIYLASTLASGVPSRFSGSGSGAEFTLTISSLQPDDFATYYCQNVYLASTNGANFGQGTKLTVLGGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCTASGFSLTDYYYMTWVRQAPGKGLEWVGFIDPDDDPYYATWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGGDHNSGWGLDIW GQGTLVTVSS

Example 3: A Tetravalent Bispecific Antibody Comprising Antibody HC2:LC1and Ranibizumab-Derived Sequences

To generate a tetravalent bispecific antibody in which the heavy chainof antibody HC2:LC1 was fused to a ranibizumab-derived VEGF-bindingdomain, an amino acid sequence was generated comprising the followingappended amino acid sequences, from N-terminus to C-terminus:

1) SEQ ID NO: 14 (heavy chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 28 (ranibizumab-derived sequence).

The resulting polypeptide (SEQ ID NO: 151) was co-expressed with SEQ IDNO: 17, to provide tetravalent, bispecific antibody HC2-RAN:LC1,comprising the sequences shown in TABLE 35. Amino acids 1-19 of SEQ IDNO: 151 are the heavy chain signal peptide (SEQ ID NO: 11). Amino acids1-20 of SEQ ID NO: 17 are the light chain signal peptide (SEQ ID NO:12). In some embodiments, a mature HC2-RAN:LC1 does not comprise thesignal peptides. For example, a mature HC2-RAN:LC1 of the disclosure cancomprise SEQ ID NO: 256 and SEQ ID NO: 250.

TABLE 35 SEQ ID NO: Name Amino acid sequence 151 SignalMGWTLVFLFLLSVTAGVHSEVQLVESGGGLVQPGGSLRLSC peptide-AASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATY HC2-YAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRD RANYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKGGGGSEVQLVESGGGLVQPGGSLRLSCAASGYDFTHYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPYYYGTSHWYFDVWGQGTLVTVSSGGGGSGGGGSGGGGSDIQLTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSLQPEDFAT YYCQQYSTVPWTFGQGTKVEIK 17Signal MVSSAQFLGLLLLCFQGTRCDVVMTQSPSFLSASVGDRVTIT peptide-CKASQHVGTAVAWYQQRPGKAPKLLIYWASTRHTGVPSRF LC1SGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGEC 256HC2- EVQLVESGGGLVQPGGSLRLSCAASGFTFNANAMNWVRQA RANPGKGLEWVGRIRTKSNNYATYYAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRDYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKGGGGSEVQLVESGGGLVQPGGSLRLSCAASGYDFTHYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPYYYGTSHWYFDVWGQGTLVTVSSGGGGSGGGGSGGGGSDIQLTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSTVPWTFGQGT KVEIK

Example 4: Characterization of Bispecific Compounds

The following multi-specific antibodies of the disclosure weregenerated: (i) HC2-RAN:LC1, (ii) HC2-AFL:LC1, (iii) HC2-BRO:LC1, (iv)HC2-ABI:LC1, (v) HC2:LC1-AFL, (vi) HC2:LC1-BRO, (vii) HC2:LC1-ABI,(viii) HC2-AFL:LC1-AFL, (ix) HC2-BRO:LC1-BRO, and (x) HC2-ABI:LC1-ABI.Enzyme-linked immunosorbent assays (ELISAs) were performed to determinebinding of these antibodies to VEGF and HPTP-β (VE-PTP). Binding toHPTP-β (VE-PTP) was confirmed for all constructs, and binding to VEGFwas confirmed for all constructs except for HC2-RAN:LC1.

The tetravalent, bispecific antibodies described in EXAMPLES 1-3 wereproduced and characterized as described in TABLE 36, FIG. 16 , FIG. 17 ,and FIG. 18 .

TABLE 36 provides results from small scale production andcharacterization of bispecific candidates.

TABLE 36 Candidate Yield HC MW LC MW Intact Name (mg) (Da)** (Da)** MW(Da) HC2-RAN:LC1 0.84 76113/76109 23400/23399 198981 HC2-AFL:LC1 1.3272756/72748 23400/23399 192270 HC2-BRO:LC1 0.73 75882/75877 23400/23399198532 **m/c = measured/calculated

FIG. 16 provides ELISA data demonstrating binding of a tetravalentbispecific antibody HC2-AFL:LC1 to HPTP-β (top panel) and VEGF (bottompanel). Binding is compared to controls (HC2:LC1 in top panel,VEGF-R2-Fc chimera in bottom panel).

FIG. 17 provides ELISA data demonstrating binding of a tetravalentbispecific antibody HC2-BRO:LC1 to HPTP-β (top panel) and VEGF (bottompanel). Binding is compared to controls (HC2:LC1 in top panel,VEGF-R2-Fc chimera in bottom panel).

FIG. 18 provides ELISA data evaluating binding of a tetravalentbispecific antibody HC2-RAN:LC1 to HPTP-β (top panel) and VEGF (bottompanel). Binding is compared to controls (HC2:LC1 in top panel,VEGF-R2-Fc chimera in bottom panel).

Example 5: A Tetravalent Bispecific Antibody Comprising Antibody HC2:LC1and Abicipar-Derived Sequences

To generate a tetravalent bispecific antibody in which the heavy chainof antibody HC2:LC1 was fused to an abicipar-derived VEGF-bindingdomain, an amino acid sequence was generated comprising the followingappended amino acid sequences, from N-terminus to C-terminus:

1) Residues 1-467 of SEQ ID NO: 14 (heavy chain of antibody HC2:LC1);

2) SEQ ID NO:31 (linker peptide, underlined); and

3) SEQ ID NO: 244 (abicipar-derived sequence).

The resulting polypeptide (SEQ ID NO: 231) was co-expressed with SEQ IDNO: 17, to provide tetravalent, bispecific antibody HC2-ABI:LC1,comprising the sequences shown in TABLE 37. Amino acids 1-19 of SEQ IDNO: 231 are the heavy chain signal peptide (SEQ ID NO: 11). Amino acids1-20 of SEQ ID NO: 17 are the light chain signal peptide (SEQ ID NO:12). In some embodiments, a mature HC2-ABI:LC1 does not comprise thesignal peptides. For example, a mature HC2-ABI:LC1 of the disclosure cancomprise SEQ ID NO: 257 and SEQ ID NO: 250.

TABLE 37 SEQ ID NO: Name Amino acid sequence 231 SignalMGWTLVFLFLLSVTAGVHSEVQLVESGGGLVQPGGSLRLSC peptide-AASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATY HC2-YAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRD ABIYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSDLDKKLLEAARAGQDDEVRILMANGADVNARDSTGWTPLHLAAPWGHPEIVEVLLKNGADVNAADFQGWTPLHLAAAVGHLEIVEVLLKYGADVNAQ DKFGKTAFDISIDNGNEDLAEILQKAA 17Signal MVSSAQFLGLLLLCFQGTRCDVVMTQSPSFLSASVGDRVTIT peptide-CKASQHVGTAVAWYQQRPGKAPKLLIYWASTRHTGVPSRF LC1SGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGEC 257HC2- EVQLVESGGGLVQPGGSLRLSCAASGFTENANAMNWVRQA ABIPGKGLEWVGRIRTKSNNYATYYAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRDYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSDLDKKLLEAARAGQDDEVRILMANGADVNARDSTGWTPLHLAAPWGHPEIVEVLLKNGADVNAADFQGWTPLHLAAAVGHLEIVEVLLKYGADVNAQDKFGKTAFDISIDNGNEDLA EILQKAA

Example 6: A Hexavalent Bispecific Antibody Comprising Antibody HC2:LC1and Brolucizumab-Derived Sequences

To generate a heavy chain with brolucizumab-derived VEGF-binding domainsadded at the C-terminus, an amino acid sequence was generated comprisingthe following appended amino acid sequences, from N-terminus toC-terminus:

1) SEQ ID NO: 14 (heavy chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 23 (brolucizumab-derived sequence).

To generate a light chain with brolucizumab-derived VEGF-binding domainsadded at the C-terminus, an amino acid sequence was generated comprisingthe following appended amino acid sequences, from N-terminus toC-terminus:

1) SEQ ID NO: 17 (light chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 23 (brolucizumab-derived sequence).

The resulting polypeptides, SEQ ID NO: 150 and SEQ ID NO: 218 wereco-expressed to provide a hexavalent, bispecific antibodyHC2-BRO:LC1-BRO shown in TABLE 38. Amino acids 1-19 of SEQ ID NO: 150are the heavy chain signal peptide (SEQ ID NO: 11). Amino acids 1-20 ofSEQ ID NO: 218 are the light chain signal peptide (SEQ ID NO: 12). Insome embodiments, a mature HC2-BRO:LC1-BRO does not comprise the signalpeptides. For example, a mature HC2-BRO:LC1-BRO of the disclosure cancomprise SEQ ID NO: 255 and SEQ ID NO: 258.

TABLE 38 SEQ ID NO: Name Amino acid sequence 150 SignalMGWTLVFLFLLSVTAGVHSEVQLVESGGGLVQPGGSLRLSC peptide-AASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATY HC2-YAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRD BROYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKGGGGSEIVMTQSPSTLSASVGDRVIITCQASEIIHSWLAWYQQKPGKAPKLLIYLASTLASGVPSRFSGSGSGAEFTLTISSLQPDDFATYYCQNVYLASTNGANFGQGTKLTVLGGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCTASGFSLTDYYYMTWVRQAPGKGLEWVGFIDPDDDPYYATWAKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCAGGDHNSGWGLDIWGQGTLVTVSS218 Signal MVSSAQFLGLLLLCFQGTRCDVVMTQSPSFLSASVGDRVTIT peptide-CKASQHVGTAVAWYQQRPGKAPKLLIYWASTRHTGVPSRF LC1-SGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEI BROKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSEIVMTQSPSTLSASVGDRVIITCQASEIIHSWLAWYQQKPGKAPKLLIYLASTLASGVPSRFSGSGSGAEFTLTISSLQPDDFATYYCQNVYLASTNGANFGQGTKLTVLGGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCTASGFSLTDYYYMTWVRQAPGKGLEWVGFIDPDDDPYYATWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGGDHNSGWGLDIWGQGTLVTVSS 258 LC1-DVVMTQSPSFLSASVGDRVTITCKASQHVGTAVAWYQQRP BROGKAPKLLIYWASTRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSEIVMTQSPSTLSASVGDRVIITCQASEIIHSWLAWYQQKPGKAPKLLIYLASTLASGVPSRFSGSGSGAEFTLTISSLQPDDFATYYCQNVYLASTNGANFGQGTKLTVLGGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCTASGFSLTDYYYMTWVRQAPGKGLEWVGFIDPDDDPYYATWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGGDHN SGWGLDIWGQGTLVTVSS

Example 7: A Hexavalent Bispecifc Antibody Comprising Antibody HC2:LC1and Aflibercept-Derived Sequences

To generate a heavy chain with aflibercept-derived VEGF-binding domainsadded at the C-terminus, an amino acid sequence was generated comprisingthe following appended amino acid sequences, from N-terminus toC-terminus:

1) SEQ ID NO: 14 (heavy chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 22 (aflibercept-derived sequence).

To generate a light chain with aflibercept-derived VEGF-binding domainsadded at the C-terminus, an amino acid sequence was generated comprisingthe following appended amino acid sequences, from N-terminus toC-terminus:

1) SEQ ID NO: 17 (light chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 22 (aflibercept-derived sequence).

The resulting polypeptides, SEQ ID NO: 149 and SEQ ID NO: 219 wereco-expressed to provide a hexavalent, bispecific antibodyHC2-AFL:LC1-AFL shown in TABLE 39. Amino acids 1-19 of SEQ ID NO: 149are the heavy chain signal peptide (SEQ ID NO: 11). Amino acids 1-20 ofSEQ ID NO: 219 are the light chain signal peptide (SEQ ID NO: 12). Insome embodiments, a mature HC2-AFL:LC1-AFL does not comprise the signalpeptides. For example, a mature HC2-AFL:LC1-AFL of the disclosure cancomprise SEQ ID NO: 254 and SEQ ID NO: 259.

TABLE 39 SEQ ID NO: Name Amino acid sequence 149 SignalMGWTLVFLFLLSVTAGVHSEVQLVESGGGLVQPGGSLRLSC peptide-AASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATY HC2-YAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRD AFLYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKGGGGSSDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAAS SGLMTKKNSTFVRVHEK 219 SignalMVSSAQFLGLLLLCFQGTRCDVVMTQSPSFLSASVGDRVTIT peptide-CKASQHVGTAVAWYQQRPGKAPKLLIYWASTRHTGVPSRF LC1-SGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEI AFLKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSSDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSD QGLYTCAASSGLMTKKNSTFVRVHEK 259LC1- DVVMTQSPSFLSASVGDRVTITCKASQHVGTAVAWYQQRP AFLGKAPKLLIYWASTRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSSDTGRPFVEMYSEIPEIMMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNST FVRVHEK

Example 8: A Hexavalent Bispecific Antibody Comprising Antibody HC2:LC1and Ranibizumab-Derived Sequences

To generate a heavy chain with ranibizumab-derived VEGF-binding domainsadded at the C-terminus, an amino acid sequence is generated comprisingthe following appended amino acid sequences, from N-terminus toC-terminus:

1) SEQ ID NO: 14 (heavy chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 28 (ranibizumab-derived sequence).

To generate a light chain with ranibizumab-derived VEGF-binding domainsadded at the C-terminus, an amino acid sequence is generated comprisingthe following appended amino acid sequences, from N-terminus toC-terminus:

1) SEQ ID NO: 17 (light chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 28 (ranibizumab-derived sequence).

The resulting polypeptides, SEQ ID NO: 151 and SEQ ID NO: 243 areco-expressed to provide a hexavalent, bispecific antibodyHC2-RAN:LC1-RAN shown in TABLE 40. Amino acids 1-19 of SEQ ID NO: 151are the heavy chain signal peptide (SEQ ID NO: 11). Amino acids 1-20 ofSEQ ID NO: 243 are the light chain signal peptide (SEQ ID NO: 12). Insome embodiments, a mature HC2-RAN:LC1-RAN does not comprise the signalpeptides. For example, a mature HC2-RAN:LC1-RAN of the disclosure cancomprise SEQ ID NO: 256 and SEQ ID NO: 260.

TABLE 40 SEQ ID NO: Name Amino acid sequence 151 SignalMGWTLVFLFLLSVTAGVHSEVQLVESGGGLVQPGGSLRLSC peptide-AASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATY HC2-YAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRD RANYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKGGGGSEVQLVESGGGLVQPGGSLRLSCAASGYDFTHYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPYYYGTSHWYFDVWGQGTLVTVSSGGGGSGGGGSGGGGSDIQLTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSLQPEDFAT YYCQQYSTVPWTFGQGTKVEIK 243Signal MVSSAQFLGLLLLCFQGTRCDVVMTQSPSFLSASVGDRVTIT peptide-CKASQHVGTAVAWYQQRPGKAPKLLIYWASTRHTGVPSRF LC1-SGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEI RANKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSEVQLVESGGGLVQPGGSLRLSCAASGYDFTHYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPYYYGTSHWYFDVWGQGTLVTVSSGGGGSGGGGSGGGGSDIQLTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSTVPWTFGQGTKVEIK 260 LC1-DVVMTQSPSFLSASVGDRVTITCKASQHVGTAVAWYQQRP RANGKAPKLLIYWASTRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSEVQLVESGGGLVQPGGSLRLSCAASGYDFTHYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPYYYGTSHWYFDVWGQGTLVTVSSGGGGSGGGGSGGGGSDIQLTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSTVP WTFGQGTKVEIK

Example 9: A Hexavalent Bispecific Antibody Comprising Antibody HC2:LC1and Abicipar-Derived Sequences

To generate a heavy chain with abicipar-derived VEGF-binding domainsadded at the C-terminus, an amino acid sequence was generated comprisingthe following appended amino acid sequences, from N-terminus toC-terminus:

1) Residues 1-467 of SEQ ID NO: 14 (heavy chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 244 (abicipar-derived sequence).

To generate a light chain with abicipar-derived VEGF-binding domainsadded at the C-terminus, an amino acid sequence was generated comprisingthe following appended amino acid sequences, from N-terminus toC-terminus:

1) SEQ ID NO: 17 (light chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 244 (abicipar-derived sequence).

The resulting polypeptides, SEQ ID NO: 231 and SEQ ID NO: 232, wereco-expressed to provide a hexavalent, bispecific antibodyHC2-ABI:LC1-ABI shown in TABLE 41. Amino acids 1-19 of SEQ ID NO: 231are the heavy chain signal peptide (SEQ ID NO: 11). Amino acids 1-20 ofSEQ ID NO: 232 are the light chain signal peptide (SEQ ID NO: 12). Insome embodiments, a mature HC2-ABI:LC1-ABI does not comprise the signalpeptides. For example, a mature HC2-ABI:LC1-ABI of the disclosure cancomprise SEQ ID NO: 257 and SEQ ID NO: 261.

TABLE 41 SEQ ID NO: Name Amino acid sequence 231 SignalMGWTLVFLFLLSVTAGVHSEVQLVESGGGLVQPGGSLRLSC peptide-AASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATY HC2-YAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRD ABIYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSDLDKKLLEAARAGQDDEVRILMANGADVNARDSTGWTPLHLAAPWGHPEIVEVLLKNGADVNAADFQGWTPLHLAAAVGHLEIVEVLLKYGADVNAQ DKFGKTAFDISIDNGNEDLAEILQKAA 232Signal MVSSAQFLGLLLLCFQGTRCDVVMTQSPSFLSASVGDRVTIT peptide-CKASQHVGTAVAWYQQRPGKAPKLLIYWASTRHTGVPSRF LC1-SGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEI ABIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSDLDKKLLEAARAGQDDEVRILMANGADVNARDSTGWTPLHLAAPWGHPEIVEVLLKNGADVNAADFQGWTPLHLAAAVGHLEIVEVLLKYGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA 261 LC1-DVVMTQSPSFLSASVGDRVTITCKASQHVGTAVAWYQQRP ABIGKAPKLLIYWASTRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSDLDKKLLEAARAGQDDEVRILMANGADVNARDSTGWTPLHLAAPWGHPEIVEVLLKNGADVNAADFQGWTPLHLAAAVGHLEIVEVLLKYGADVNAQDKFGKTAFDISI DNGNEDLAEILQKAA

Example 10: A Tetravalent Bispecific Antibody Comprising AntibodyHC2:LC1 and Brolucizumab-Derived Sequences

To generate a light chain with brolucizumab-derived VEGF-binding domainsadded at the C-terminus, an amino acid sequence was generated comprisingthe following appended amino acid sequences, from N-terminus toC-terminus:

1) SEQ ID NO: 17 (light chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 23 (brolucizumab-derived sequence).

The resulting polypeptide (SEQ ID NO: 218) was co-expressed with SEQ IDNO: 14, to provide tetravalent, bispecific antibody HC2:LC1-BRO,comprising the sequences shown in TABLE 42. Amino acids 1-19 of SEQ IDNO: 14 are the heavy chain signal peptide (SEQ ID NO: 11). Amino acids1-20 of SEQ ID NO: 218 are the light chain signal peptide (SEQ ID NO:12). In some embodiments, a mature HC2:LC1-BRO does not comprise thesignal peptides. For example, a mature HC2:LC1-BRO of the disclosure cancomprise SEQ ID NO: 247 and SEQ ID NO: 258.

TABLE 42 SEQ ID NO: Name Amino acid sequence 14 SignalMGWTLVFLFLLSVTAGVHSEVQLVESGGGLVQPGGSLRLSC peptide-AASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATY HC2YAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRDYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMH EALHNHYTQKSLSLSLGK 218 SignalMVSSAQFLGLLLLCFQGTRCDVVMTQSPSFLSASVGDRVTIT peptide-CKASQHVGTAVAWYQQRPGKAPKLLIYWASTRHTGVPSRF LC1-SGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEI BROKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSEIVMTQSPSTLSASVGDRVIITCQASEIIHSWLAWYQQKPGKAPKWYLASTLASGVPSRFSGSGSGAEFTLTISSLQPDDFATYYCQNVYLASTNGANFGQGTKLTVLGGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCTASGFSLTDYYYMTWVRQAPGKGLEWVGFIDPDDDPYYATWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGGDHNSGWGLDIWGQGTLVTVSS

Example 11: A Tetravalent Bispecific Antibody Comprising AntibodyHC2:LC1 and Aflibercept-Derived Sequences

To generate a light chain with aflibercept-derived VEGF-binding domainsadded at the C-terminus, an amino acid sequence was generated comprisingthe following appended amino acid sequences, from N-terminus toC-terminus:

1) SEQ ID NO: 17 (light chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 22 (aflibercept-derived sequence).

The resulting polypeptide (SEQ ID NO: 219) was co-expressed with SEQ IDNO: 14, to provide tetravalent, bispecific antibody HC2:LC1-AFL,comprising the sequences shown in TABLE 43. Amino acids 1-19 of SEQ IDNO: 14 are the heavy chain signal peptide (SEQ ID NO: 11). Amino acids1-20 of SEQ ID NO: 219 are the light chain signal peptide (SEQ ID NO:12). In some embodiments, a mature HC2:LC1-AFL does not comprise thesignal peptides. For example, a mature HC2:LC1-AFL of the disclosure cancomprise SEQ ID NO: 247 and SEQ ID NO: 259.

TABLE 43 SEQ ID NO: Name Amino acid sequence 14 SignalMGWTLVFLFLLSVTAGVHSEVQLVESGGGLVQPGGSLRLSC peptide-AASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATY HC2YAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRDYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMH EALHNHYTQKSLSLSLGK 219 SignalMVSSAQFLGLLLLCFQGTRCDVVMTQSPSFLSASVGDRVTIT peptide-CKASQHVGTAVAWYQQRPGKAPKLLIYWASTRHTGVPSRF LC1-SGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEI AFLKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSSDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSD QGLYTCAASSGLMTKKNSTFVRVHEK

Example 12: A Tetravalent Bispecific Antibody Comprising AntibodyHC2:LC1 and Ranibizumab-Derived Sequences

To generate a light chain with ranibizumab-derived VEGF-binding domainsadded at the C-terminus, an amino acid sequence is generated comprisingthe following appended amino acid sequences, from N-terminus toC-terminus:

1) SEQ ID NO: 17 (light chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 28 (ranibizumab-derived sequence).

The resulting polypeptide (SEQ ID NO: 243) is co-expressed with SEQ IDNO: 14, to provide tetravalent, bispecific antibody HC2:LC1-RAN,comprising the sequences shown in TABLE 44. Amino acids 1-19 of SEQ IDNO: 14 are the heavy chain signal peptide (SEQ ID NO: 11). Amino acids1-20 of SEQ ID NO: 243 are the light chain signal peptide (SEQ ID NO:12). In some embodiments, a mature HC2:LC1-RAN does not comprise thesignal peptides. For example, a mature HC2:LC1-RAN of the disclosure cancomprise SEQ ID NO: 247 and SEQ ID NO: 260.

TABLE 44 SEQ ID NO: Name Amino acid sequence 14 SignalMGWTLVFLFLLSVTAGVHSEVQLVESGGGLVQPGGSLRLSC peptide-AASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATY HC2YAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRDYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMH EALHNHYTQKSLSLSLGK 243 SignalMVSSAQFLGLLLLCFQGTRCDVVMTQSPSFLSASVGDRVTIT peptide-CKASQHVGTAVAWYQQRPGKAPKLLIYWASTRHTGVPSRF LC1-SGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEI RANKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSEVQLVESGGGLVQPGGSLRLSCAASGYDFTHYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPYYYGTSHWYFDVWGQGTLVTVSSGGGGSGGGGSGGGGSDIQLTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSTVPWTFGQGTKVEIK

Example 13: A Tetravalent Bispecific Antibody Comprising AntibodyHC2:LC1 and Abicipar-Derived Sequences

To generate a light chain with abicipar-derived VEGF-binding domainsadded at the C-terminus, an amino acid sequence was generated comprisingthe following appended amino acid sequences, from N-terminus toC-terminus:

1) SEQ ID NO: 17 (light chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 244 (abicipar-derived sequence).

The resulting polypeptide (SEQ ID NO: 232) was co-expressed with SEQ IDNO: 245 (residues 1-467 of SEQ ID NO: 14), to provide tetravalent,bispecific antibody HC2:LC1-ABI, comprising the sequences shown in TABLE45. Amino acids 1-19 of SEQ ID NO: 245 are the heavy chain signalpeptide (SEQ ID NO: 11). Amino acids 1-20 of SEQ ID NO: 232 are thelight chain signal peptide (SEQ ID NO: 12). In some embodiments, amature HC2:LC1-ABI does not comprise the signal peptides. For example, amature HC2:LC1-ABI of the disclosure can comprise SEQ ID NO: 247 and SEQID NO: 261.

TABLE 45 SEQ ID NO: Name Amino acid sequence 245 SignalMGWTLVFLFLLSVTAGVHSEVQLVESGGGLVQPGGSLRLSC peptide-AASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATY HC2YAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRDYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMH EALHNHYTQKSLSLSLG 232 SignalMVSSAQFLGLLLLCFQGTRCDVVMTQSPSFLSASVGDRVTIT peptide-CKASQHVGTAVAWYQQRPGKAPKLLIYWASTRHTGVPSRF LC1-SGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEI ABIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSDLDKKLLEAARAGQDDEVRILMANGADVNARDSTGWTPLHLAAPWGHPEIVEVLLKNGADVNAADFQGWTPLHLAAAVGHLEIVEVLLKYGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA 262 HC2EVQLVESGGGLVQPGGSLRLSCAASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATYYAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRDYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG

Example 14: Bispecific Antibodies that Activate Tie2

Human Umbilical Cord Endothelial Cells (HUVECs) were seeded onto T75flasks coated with porcine gelatin. Cell maintenance was performed usingcomplete medium (EGM or EGM-2) and sub-cultured using Trypsin/EDTA into100 mm dishes. After 3 days, the 100 mm dishes were rinsed, and treatedin basal medium (EBM, EBM-2, OptiMEM I) for 30 minutes at 37° C./5% CO₂with one of the following antibodies at 5 or 50 nM as indicated in FIG.19 : (i) HC2:LC1, a humanized monoclonal antibody specific for HPTP-β;(ii) HC2-BRO:LC1, a tetravalent bispecific antibody comprisingbrolucizumab-derived VEGF-binding domains fused to the C-termini of theheavy chains of HC2:LC1; (iii) HC2-AFL:LC1, a tetravalent bispecificantibody comprising aflibercept-derived VEGF-binding domains fused tothe C-termini of the heavy chains of HC2:LC1; (iv) HC2-BRO:LC1-BRO, ahexavalent antibody comprising brolucizumab-derived VEGF-binding domainsfused to the C-termini of the heavy chains and the light chains ofHC2:LC1; or (v) HC2-AFL:LC1-AFL, a hexavalent antibody comprisingaflibercept-derived VEGF-binding domains fused to the C-termini of theheavy chains and the light chains of HC2:LC1. After treatment, the cellswere rinsed with ice cold PBS containing 1 mM NaOV and lysed in CompleteTriton X Lysis Buffer (20 mM Tris-HCl, 137 mM NaCl, 10% Glycerol, 1%Triton X-100, 2 mM EDTA, 1 mM NaOV, 1 mM NaF, 1 mM PMSF, 1 μg/mLleupeptin, 1 μg/mL pepstatin).

The lysates were immunoprecipitated with anti-Tie2 and anti-VEGFR2antibodies. 1-10 μg of VEGFR2 antibody (MAB3573, #89109), TIE-2 antibody(Ab33), and 25 μL of Protein A/G agarose beads were added to 1 mL ofHUVEC lysate, and the tubes were placed on a rotating platform for 1-3days at 4° C. The IP reaction tubes were rinsed with 1 mL completeTriton-X lysis buffer and resuspended in 2× loading dye containing DTT.The tubes were incubated at 95° C. for five minutes, spun down, and 25μL loaded per well into a Tris Glycine gel. The samples were resolved ona gel, transferred to a PVDF membrane, and subjected to serial westernblot to detect phosphotyrosine (phospho-Tie2 and phospho-VEGFR2),followed by re-probing to blot for total Tie2, and total VEGFR2. The gelwas run at 125V for 75 minutes, before transfer to a PVDF membrane. Themembranes were blocked in 5% BSA/0.05% Tween Tris Wash Buffer for 1 hourat room temperature (RT) on a rotating platform. Primary antibodies(PY99; VEGFR2-A3; TIE-2 Ab33) were added for 1 hour at a 1:1000 dilutionin wash buffer. Secondary antibody (anti-mouse hrp) was added for 1 hourat a 1:1000 dilution in wash buffer. The membranes were rinsed threetimes with 0.05-0.1% Tween 20+Tris Buffered Saline (TBS) between steps.An ECL detection system was used to visualize bands. Re-probe wasperformed on lots treated with 200 mM Glycine for 24-48 h.

As shown in FIG. 19 , all of the tested bispecific antibodies increasedbasal Tie2 activation in HUVECS, while basal VEGFR2 phosphorylation wasnot affected. The top panel provides a western blot showing Tie2activation and VEGFR2 activation as shown through detection ofphospho-Tie2 and phospho-VEGFR2, respectively (top half), and total Tie2and VEGFR2 (bottom half). The bottom panel provides the densitometricratios of phosphorylated Tie2 to total Tie2.

The assay was repeated with the following antibodies of the disclosure,which also enhanced Tie2 activation in the absence of exogenous Ang1:(i) HC2-RAN:LC1, (ii) HC2-ABI:LC1, (iii) HC2:LC1-AFL, (iv) HC2:LC1-BRO,(v) HC2:LC1-ABI, and (vi) HC2-ABI:LC1-ABI.

Example 15: Bispecific Antibodies Enhance Ang1-Mediated Tie2 Activationand Block VEGF-Mediated VEGFR2 Activation

Human Umbilical Cord Endothelial Cells (HUVECs) were seeded onto T75flasks coated with porcine gelatin. Cell maintenance was performed usingcomplete medium (EGM or EGM-2) and sub-cultured using Trypsin/EDTA into100 mm dishes. After 3 days, the 100 mm dishes were rinsed, andmock-pre-treated or pre-treated in basal medium (EBM, EBM-2, OptiMEM I)for 30 minutes at 37° C./5% CO₂ with one of the following antibodies at5 or 50 nM as indicated in FIG. 20 : (i) HC2:LC1, a humanized monoclonalantibody specific for HPTP-β; (ii) HC2-BRO:LC1, a tetravalent bispecificantibody comprising brolucizumab-derived VEGF-binding domains fused tothe C-termini of the heavy chains of HC2:LC1; (iii) HC2-AFL:LC1, atetravalent bispecific antibody comprising aflibercept-derivedVEGF-binding domains fused to the C-termini of the heavy chains ofHC2:LC1; (iv) HC2-BRO:LC1-BRO, a hexavalent antibody comprisingbrolucizumab-derived VEGF-binding domains fused to the C-termini of theheavy chains and the light chains of HC2:LC1; or (v) HC2-AFL:LC1-AFL, ahexavalent antibody comprising aflibercept-derived VEGF-binding domainsfused to the C-termini of the heavy chains and the light chains ofHC2:LC1. After pre-treatment, cells were treated with VEGF (5 ng/mL) andAng1 (50 ng/mL) for 6 minutes at 37° C./5% CO₂ in basal medium(Phosphate buffered saline, PBS+0.2% Bovine Serum Albumin, or OptiMEMI). After treatment, the cells were rinsed with ice cold PBS containing1 mM NaOV and lysed in Complete Triton X Lysis Buffer (20 mM Tris-HCl,137 mM NaCl, 10% Glycerol, 1% Triton X-100, 2 mM EDTA, 1 mM NaOV, 1 mMNaF, 1 mM PMSF, 1 μg/mL leupeptin, 1 μg/mL pepstatin).

The lysates were immunoprecipitated with anti-Tie2 and anti-VEGFR2antibodies. 1-10 μg of VEGFR2 antibody (MAB3573, #89109), TIE-2 antibody(Ab33), and 25 μL of Protein A/G agarose beads were added to 1 mL ofHUVEC lysate, and the tubes were placed on a rotating platform for 1-3days at 4° C. The IP reaction tubes were rinsed with 1 mL completeTriton-X lysis buffer and resuspended in 2× loading dye containing DTT.The tubes were incubated at 95° C. for five minutes, spun down, and 25μL loaded per well into a Tris Glycine gel. The samples were resolved ona gel, transferred to a PVDF membrane, and subjected to serial westernblot to detect phosphotyrosine (phospho-Tie2 and phospho-VEGFR2),followed by re-probing to blot for total Tie2, and total VEGFR2. The gelwas run at 125V for 75 minutes, before transfer to a PVDF membrane. Themembranes were blocked in 5% BSA/0.05% Tween Tris Wash Buffer for 1 hourat room temperature (RT) on a rotating platform. Primary antibodies(PY99; VEGFR2-A3; TIE-2 Ab33) were added for 1 hour at a 1:1000 dilutionin wash buffer. Secondary antibody (anti-mouse hrp) was added for 1 hourat a 1:1000 dilution in wash buffer. The membranes were rinsed threetimes with 0.05-0.1% Tween 20+Tris Buffered Saline (TBS) between steps.An ECL detection system was used to visualize bands. Re-probe wasperformed on lots treated with 200 mM Glycine for 24-48 h.

Treatment with VEGF and Ang1 resulted in increased phosphorylation ofVEGFR2 and Tie2 (FIG. 20 ). Treatment with the HC2:LC1 antibody,specific for HPTP-β, enhanced Ang1-mediated Tie2 activation in cellstreated with Ang1 and VEGF. Treatment with the bispecific antibodiesenhanced Ang1-mediated Tie2 activation and blocked VEGF-mediated VEGFR2activation in cells treated with Ang1 and VEGF. The top panel provides awestern blot showing Tie2 activation and VEGFR2 activation as shownthrough detection of phospho-Tie2 and phospho-VEGFR2, respectively (tophalf), and total Tie2 and VEGFR2 (bottom half). The lower panel providesthe densitometric ratios of phosphorylated Tie2 to total Tie2 andphosphorylated VEGFR2 to total VEGFR2.

Example 16: Bispecific Antibodies Enhance Ang1-Mediated Tie2 Activationand Block VEGF-Mediated VEGFR2 Activation (Immunoprecipitation andWestern Blot Assays)

Human Umbilical Cord Endothelial Cells (HUVECs) were seeded onto T75flasks coated with porcine gelatin. Cell maintenance was performed usingcomplete medium (EGM or EGM-2) and sub-cultured using Trypsin/EDTA into100 mm dishes. After 3 days, the 100 mm dishes were rinsed, andpre-treated in basal medium (EBM, EBM-2, OptiMEM I) for 30 minutes at37° C./5% CO₂ with one of multi-specific antibodies of the disclosure asindicated for each figure. After pre-treatment, the cells weremock-treated or treated with VEGF and Ang1 for 6 minutes at 37° C./5%CO₂ in basal medium (Phosphate buffered saline, PBS+0.2% Bovine SerumAlbumin, or OptiMEM I). After treatment, the cells were rinsed with icecold PBS containing 1 mM NaOV and lysed in Complete Triton X LysisBuffer (20 mM Tris-HCl, 137 mM NaCl, 10% Glycerol, 1% Triton X-100, 2 mMEDTA, 1 mM NaOV, 1 mM NaF, 1 mM PMSF, 1 μg/mL leupeptin, 1 μg/mLpepstatin).

The lysates were immunoprecipitated with anti-Tie2 and anti-VEGFR2antibodies. 1-10 g of VEGFR2 antibody (MAB3573, #89109), TIE-2 antibody(Ab33), and 25 μL of Protein A/G agarose beads were added to 1 mL ofHUVEC lysate, and the tubes were placed on a rotating platform for 1-3days at 4° C. The IP reaction tubes were rinsed with 1 mL completeTriton-X lysis buffer and resuspended in 2× loading dye containing DTT.The tubes were incubated at 95° C. for five minutes, spun down, and 25μL loaded per well into a Tris Glycine gel. The samples were resolved ona gel, transferred to a PVDF membrane, and subjected to serial westernblot to detect phosphotyrosine (phospho-Tie2 and phospho-VEGFR2),followed by re-probing to blot for total Tie2, and total VEGFR2. The gelwas run at 125V for 75 minutes, before transfer to a PVDF membrane. Themembranes were blocked in 5% BSA/0.05% Tween Tris Wash Buffer for 1 hourat room temperature (RT) on a rotating platform. Primary antibodies(PY99; VEGFR2-A3; TIE-2 Ab33) were added for 1 hour at a 1:1000 dilutionin wash buffer. Secondary antibody (anti-mouse hrp) was added for 1 hourat a 1:1000 dilution in wash buffer. The membranes were rinsed threetimes with 0.05-0.1% Tween 20+Tris Buffered Saline (TBS) between steps.An ECL detection system was used to visualize bands. Re-probe wasperformed on lots treated with 200 mM Glycine for 24-48 h. The assay wasrepeated using different multi-specific antibodies of the disclosure,and different concentrations of VEGF and Ang1 as indicated for thefollowing figures.

For FIG. 21A, FIG. 21B, and FIG. 21C, the cells were mock-pre-treated orpre-treated at 5 nM or 50 nM with one of: (i) HC2-ABI:LC1, a tetravalentbispecific antibody comprising abicipar-derived VEGF-binding domainsfused to the C-termini of the heavy chains of HC2:LC1 (a humanizedmonoclonal antibody specific for HPTP-β); (ii) HC2-ABI:LC1-ABI, ahexavalent antibody comprising abicipar-derived VEGF-binding domainsfused to the C-termini of the heavy chains and the light chains ofHC2:LC1; or (iii) HC2-AFL:LC1-AFL, a hexavalent antibody comprisingaflibercept-derived VEGF-binding domains fused to the C-termini of theheavy chains and the light chains of HC2:LC1. After pre-treatment, thecells were mock-treated (−) or treated (+) with VEGF (5 ng/mL) and Ang1(50 ng/mL). Treatment with VEGF and Ang1 resulted in increasedphosphorylation of VEGFR2 and Tie2 (FIG. 21A, FIG. 21B, and FIG. 21C).Treatment with the bispecific antibodies enhanced Tie2 activation andblocked VEGFR2 activation, including in cells treated with Ang1 and VEGF(FIG. 21A, FIG. 21B, and FIG. 21C). FIG. 21A provides a western blotshowing Tie2 activation and VEGFR2 activation as shown through detectionof phospho-Tie2 and phospho-VEGFR2, respectively (top half), and totalTie2 and VEGFR2 (bottom half). FIG. 21B provides the densitometric ratioof phosphorylated Tie2 to total Tie2, normalized to untreated cells.FIG. 21C provides the densitometric ratio of phosphorylated VEGFR2 tototal VEGFR2, normalized to untreated cells.

For FIG. 22A, FIG. 22B, and FIG. 22C, the cells were mock-pre-treated orpre-treated with one of the following antibodies at 5 nM: (i)HC2-BRO:LC1-BRO, a hexavalent antibody comprising brolucizumab-derivedVEGF-binding domains fused to the C-termini of the heavy chains and thelight chains of HC2:LC1 (a humanized monoclonal antibody specific forHPTP-β); (ii) HC2-AFL:LC1-AFL, a hexavalent antibody comprisingaflibercept-derived VEGF-binding domains fused to the C-termini of theheavy chains and the light chains of HC2:LC1; or (iii) HC2-ABI:LC1:ABI,a hexavalent bispecific antibody comprising abicipar-derivedVEGF-binding domains fused to the C-termini of the heavy chains and thelight chains of HC2:LC1. After pre-treatment, the cells weremock-treated (−), treated with 5 ng/mL VEGF and 50 ng/mL Ang1 (+), ortreated with 50 ng/mL VEGF and 250 ng/mL of Ang1 (+++). Treatment withVEGF and Ang1 resulted in increased phosphorylation of VEGFR2 and Tie2(FIG. 22A, FIG. 22B, and FIG. 22C). Treatment with the bispecificantibodies enhanced Tie2 activation and blocked VEGFR2 activation,including in cells treated with Ang1 and VEGF. (FIG. 22A, FIG. 22B, andFIG. 22C). FIG. 22A provides a western blot showing Tie2 activation andVEGFR2 activation as shown through detection of phospho-Tie2 andphospho-VEGFR2, respectively (top half), and total Tie2 and VEGFR2(bottom half). FIG. 22B provides the densitometric ratio ofphosphorylated Tie2 to total Tie2, normalized to untreated cells. FIG.22C provides the densitometric ratio of phosphorylated VEGFR2 to totalVEGFR2, normalized to untreated cells.

Example 17: Bispecific Antibodies Enhance Ang1-Mediated Tie2 Activationand Block VEGF-Mediated VEGFR2 Activation (ElectrochemiluminescenceAssays)

Human Umbilical Cord Endothelial Cells (HUVECs) were seeded onto T75flasks coated with porcine gelatin. Cell maintenance was performed usingcomplete medium (EGM or EGM-2) and sub-cultured using Trypsin/EDTA into100 mm dishes. After 3 days, the 100 mm dishes were rinsed, andmock-pre-treated or pre-treated in basal medium (EBM, EBM-2, OptiMEM I)for 30 minutes at 37° C./5% CO₂ with one the antibodies indicated belowfor each figure.

After pre-treatment, the cells were mock-treated or treated with VEGFand Ang1 for 6 minutes at 37° C./5% CO₂ in basal medium (Phosphatebuffered saline, PBS+0.2% Bovine Serum Albumin, or OptiMEM I). Aftertreatment, the cells were rinsed with ice cold PBS containing 1 mM NaOVand lysed in Complete Triton X Lysis Buffer (20 mM Tris-HCl, 137 mMNaCl, 10% Glycerol, 1% Triton X-100, 2 mM EDTA, 1 mM NaOV, 1 mM NaF, 1mM PMSF, 1 μg/mL leupeptin, 1 μg/mL pepstatin).

Phosphorylated Tie2 and phosphorylated VEGFR2 were quantified byelectrochemiluminescence. Primary or capture Antibodies were spottedonto 96 well Sector Imager plates. 5 μL of Tie2 antibody (30 μg/mL,AF313) or VEGFR2 antibody (30 μg/mL, 89109) were coated for 1 hour (atroom temperature) or overnight (4° C.). Each plate was washed threetimes with TBS+0.02% Tween 20 (this was performed between each step).Wells were blocked with MSD Blocker A-3% in wash buffer for 1 hour on arotating platform at room temperature. 25 μL of HUVEC lysates were addedto each well directly and incubated for 1 hour on a rotating platform atroom temperature. Detection antibody (1) was diluted to 2 μg/mL in 1%blocker/wash buffer (Ab33; AF2720/Y992; pTyr 1214; NB100 530), and 25μL/well incubated for 1 hour on a rotating platform at room temperature.Detection antibody (2) was diluted to 1 μg/mL in 1% blocker/wash buffer(Goat anti-mouse, GAM; Goat anti-rabbit, GAR; both Sulfo Tag-labeled)and 25 μL/well incubated for 1 hour on a rotating platform at roomtemperature. Signal was captured using 150 μL of Meso Scale Discovery(MSD) read buffer in each well on an MSD imager instrument. The assaywas repeated using different multi-specific antibodies of thedisclosure, and different concentrations of VEGF and Ang1 as indicatedfor the following figures.

For FIG. 23A and FIG. 23B, the cells were mock-treated or pre-treatedwith one of the following antibodies at 5 nM: (i) HC2-BRO:LC1, atetravalent bispecific antibody comprising brolucizumab-derivedVEGF-binding domains fused to the C termini of the heavy chains ofHC2:LC1 (a humanized monoclonal antibody specific for HPTP-β); (ii)HC2:LC1-BRO, a tetravalent bispecific antibody comprisingbrolucizumab-derived VEGF-binding domains fused to the C termini of thelight chains of HC2:LC1; (iii) HC2-BRO:LC1-BRO, a hexavalent bispecificantibody comprising brolucizumab-derived VEGF-binding domains fused tothe C termini of the heavy chains and the light chains of HC2:LC1; (iv)HC2-AFL:LC1, a tetravalent bispecific antibody comprisingaflibercept-derived VEGF-binding domains fused to the C termini of theheavy chains of HC2:LC1; (v) HC2:LC1-AFL, a tetravalent bispecificantibody comprising aflibercept-derived VEGF-binding domains fused tothe C termini of the light chains of HC2:LC1; (vi) HC2-AFL:LC1-AFL, ahexavalent bispecific antibody comprising aflibercept-derivedVEGF-binding domains fused to the C termini of the heavy chains and thelight chains of HC2:LC1; (vii) HC2-ABI:LC1, a tetravalent bispecificantibody comprising abicipar-derived VEGF-binding domains fused to the Ctermini of the heavy chains of HC2:LC1; (viii) HC2:LC1-ABI, atetravalent bispecific antibody comprising abicipar-derived VEGF-bindingdomains fused to the C termini of the light chains of HC2:LC1; or (ix)HC2-ABI:LC1-ABI, a hexavalent bispecific antibody comprisingabicipar-derived VEGF-binding domains fused to the C termini of theheavy chains and the light chains of HC2:LC1. FIG. 23A and FIG. 23Bprovide the ratio of phosphorylated Tie2 to total Tie2 andphosphorylated VEGFR2 to total VEGFR2, respectively, normalized tountreated cells. Treatments were as follows: mock-treated (−), treatedwith 5 ng/mL VEGF and 50 ng/mL Ang1 (+), or treated with 25 ng/mL VEGFand 250 ng/mL of Ang1 (+++). Treatment with VEGF and Ang1 resulted inincreased phosphorylation of Tie2 (FIG. 23A) and VEGFR2 (FIG. 23B).Pre-treatment with the multi-specific antibodies enhanced Tie2activation (FIG. 23A) and inhibited VEGFR2 activation (FIG. 23B) incells treated with Ang1 and VEGF.

For FIG. 24A and FIG. 24B, the cells were mock-treated or pre-treatedwith one of the following antibodies at 5 nM: (i) HC2-BRO:LC1-BRO, ahexaavalent bispecific antibody comprising brolucizumab-derivedVEGF-binding domains fused to the C termini of the heavy chains and thelight chains of HC2:LC1 (a humanized monoclonal antibody specific forHPTP-β); (ii) HC2-AFL:LC1-AFL, a hexavalent bispecific antibodycomprising aflibercept-derived VEGF-binding domains fused to the Ctermini of the heavy chains and the light chains of HC2:LC1; or (iii)HC2-ABI:LC1-ABI, a hexavalent bispecific antibody comprisingabicipar-derived VEGF-binding domains fused to the C termini of theheavy chains and the light chains of HC2:LC1. FIG. 24A and FIG. 24Bprovide the ratio of phosphorylated Tie2 to total Tie2 andphosphorylated VEGFR2 to total VEGFR2, respectively, normalized tountreated cells. Treatments were as follows: mock-treated (−), treatedwith 5 ng/mL VEGF and 50 ng/mL Ang1 (+), or treated with 25 ng/mL VEGFand 250 ng/mL of Ang1 (+++). Treatment with VEGF and Ang1 resulted inincreased phosphorylation of Tie2 (FIG. 24A) and VEGFR2 (FIG. 24B).Pre-treatment with the multi-specific antibodies enhanced Tie2activation (FIG. 24A) and inhibited VEGFR2 activation (FIG. 24B) incells treated with Ang1 and VEGF.

For FIG. 25A and FIG. 25B, the cells were mock-treated or pre-treatedwith one of the following antibodies at 5 or 50 nM: (i) HC2-BRO:LC1, atetravalent bispecific antibody comprising brolucizumab-derivedVEGF-binding domains fused to the C termini of the heavy chains ofHC2:LC1 (a humanized monoclonal antibody specific for HPTP-β); or (ii)HC2-BRO:LC1-BRO, a hexavalent bispecific antibody comprisingbrolucizumab-derived VEGF-binding domains fused to the C termini of theheavy chains and the light chains of HC2:LC1. FIG. 25A and FIG. 25Bprovide the ratio of phosphorylated Tie2 to total Tie2 andphosphorylated VEGFR2 to total VEGFR2, respectively, normalized tountreated cells. Treatments were as follows: mock-treated (−), treatedwith 5 ng/mL VEGF and 50 ng/mL Ang1 (+), or treated with 25 ng/mL VEGFand 250 ng/mL of Ang1 (+++). Treatment with VEGF and Ang1 resulted inincreased phosphorylation of Tie2 (FIG. 25A) and VEGFR2 (FIG. 25B).Pre-treatment with the multi-specific antibodies enhanced Tie2activation (FIG. 25A) and inhibited VEGFR2 activation (FIG. 25B) incells treated with Ang1 and VEGF.

For FIG. 26A and FIG. 26B, the cells were mock-treated or pre-treatedwith one of the following antibodies at 5 nM or 50 nM: (i) HC2-ABI:LC1,a tetravalent bispecific antibody comprising abicipar-derivedVEGF-binding domains fused to the C termini of the heavy chains ofHC2:LC1 (a humanized monoclonal antibody specific for HPTP-β); (ii)HC2-ABI:LC1-ABI, a hexavalent bispecific antibody comprisingabicipar-derived VEGF-binding domains fused to the C termini of theheavy chains and the light chains of HC2:LC1; or (iii) HC2-AFL:LC1:AFL,a hexavalent bispecific antibody comprising aflibercept-derivedVEGF-binding domains fused to the C termini of the heavy chains and thelight chains of HC2:LC1. FIG. 26A and FIG. 26B provide the ratio ofphosphorylated Tie2 to total Tie2 and phosphorylated VEGFR2 to totalVEGFR2, respectively, normalized to untreated cells. Treatments were asfollows: mock-treated (−), or treated with 5 ng/mL VEGF and 50 ng/mLAng1 (+). Treatment with VEGF and Ang1 resulted in increasedphosphorylation of Tie2 (FIG. 26A) and VEGFR2 (FIG. 26B). Pre-treatmentwith the multi-specific antibodies enhanced Tie2 activation (FIG. 26A)and inhibited VEGFR2 activation (FIG. 26B) in cells treated with Ang1and VEGF.

For FIG. 27A and FIG. 27B, the cells were mock-treated or pre-treatedwith one of the following antibodies at 5 nM or 50 nM: (i) HC2-ABI:LC1,a tetravalent bispecific antibody comprising abicipar-derivedVEGF-binding domains fused to the C termini of the heavy chains ofHC2:LC1 (a humanized monoclonal antibody specific for HPTP-β); (ii)HC2-ABI:LC1-ABI, a hexavalent bispecific antibody comprisingabicipar-derived VEGF-binding domains fused to the C termini of theheavy chains and the light chains of HC2:LC1; or (iii) HC2-AFL:LC1, atetravalent bispecific antibody comprising aflibercept-derivedVEGF-binding domains fused to the C termini of the heavy chains ofHC2:LC1. FIG. 27A and FIG. 27B provide the ratio of phosphorylated Tie2to total Tie2 and phosphorylated VEGFR2 to total VEGFR2, respectively,normalized to untreated cells. Treatments were as follows: mock-treated(−), or treated with 25 ng/mL VEGF and 250 ng/mL Ang1 (+). Treatmentwith VEGF and Ang1 resulted in increased phosphorylation of Tie2 (FIG.27A) and VEGFR2 (FIG. 27B). Pre-treatment with the multi-specificantibodies enhanced Tie2 activation (FIG. 27A) and inhibited VEGFR2activation (FIG. 27B) in cells treated with Ang1 and VEGF.

Example 18: Multi-Specific Antibodies Bind HPTP-β and VEGF at HighAffinity (Biacore Surface Plasmon Resonance Assays)

Biacore surface plasmon resonance assays were performed to determine theequilibrium dissociation constant (K_(D)) of antibodies of thedisclosure for VEGF and HPTP-β (VE-PTP). Binding experiments wereperformed on Biacore 3000/Biacore T-200 instruments at 25° C.

The assay buffer contained 10 mM HEPES buffer (pH 7.4), 150 mM NaCl, 3mM EDTA, and 0.05% P20 (polyoxyethylenesorbitan). The regenerationbuffer contained 10 mM Glycine buffer (pH 1.75). The conjugation buffercontained 10 mM sodium acetate buffer (pH 5). A flow rate of 5 μL/minutewas used for capturing ligand. A flow rate of 30 μL/minute was used forkinetic analysis.

For analysis of binding of antibodies to HPTP-β, poly-histidine taggedHPTP-β extracellular domain (ECD) 1/2 was immobilized on a chip surfacevia anti-His antibodies. Goat anti-His antibody was first immobilized onthe surface of the chip by direct immobilization using EDC/NHS(N-ethyl-N′-(3-dimethyl aminopropyl carbodiimide)/N-hydroxy succinamide)coupling chemistry on flow cell 2 of the CM5 (Carboxymethylated dextrancoated) chip. Unoccupied sites were blocked with 1M ethanolamine. TheHis-tagged ligand HPTP-β 1/2 ECD was captured at a response unit (RU) of50. The analyte (antibody) was flowed over the chip at a single analyteconcentration at a time. The binding of analyte to the ligand wasmonitored in real time to obtain on (k_(a)) and off (k_(d)) rates. Theequilibrium constant (K_(D)) was calculated from the observed k_(a) andk_(d).

For analysis of binding of the antibodies to VEGF, the antibodies wereimmobilized on the surface of the chip by direct immobilization usingEDC/Ni s coupling chemistry on flow cell 2 of the CM5 chip. Unoccupiedsites were blocked with 1M ethanolamine. The analyte (VEGF) was flowedover the chip at a single analyte concentration at a time. The bindingof analyte to the ligand was monitored in real time to obtain on (k_(a))and off (k_(d)) rates. The equilibrium constant (K_(D)) was calculatedfrom the observed k_(a) and k_(d).

Scouting analysis was performed using 10 nM of analyte to determineapproximate K_(D). Chi square analysis was carried out between theactual sensorgram and the sensorgram generated from the BIAnalysissoftware to confirm the accuracy of the analysis; values of 1-2 wereconsidered accurate and below 1 highly accurate.

TABLE 46 provides equilibrium dissociation constants based on scoutingexperiments performed at a single ligand concentration. NB=nosignificant binding detected.

TABLE 46 VEGF VEGF HPTP-β HPTP-β Construct K_(D) chi sq K_(D) chi sqHC2:LC1 NB NB 7.32E−11M 0.0439 (73.2 pM) Aflibercept 1.60E−10M 0.278 NBNB (160 pM) HC2-AFL:LC1 3.50E−10M 0.13 3.20E−10M 0.336 (350 pM) (320 pM)HC2-BRO:LC1 1.12E−13M 0.148 3.25E−10M 0.323 (112 fM) (325 pM)HC2-ABI:LC1 1.10E−13M 1.68 1.22E−10M 1.38 (110 fM) (122 pM) HC2-RBZ:LC1NB 5.86E−13M 0.16 (586 fM) HC2:LC1-AFL 8.25E−10M 0.189 4.67E−10M 1.48(825 pM) (467 pM) HC2:LC1-BRO 3.51E−14M 0.185 3.09E−10M 0.691 (35.1 fM)(309 pM) HC2:LC1-ABI 5.19E−10M 3.09 6.10E−11M 1.47 (519 pM) (61 pM)HC2-AFL:LC1-AFL 5.57E−10M 2.53 6.94E−11M 3.14 (557 pM) (69.4 pM)HC2-BRO:LC1-BRO 1.79E−12M 0.809 3.50E−12M 2.15 (1.79 pM) (3.5 pM)HC2-ABI:LC1-ABI 6.80E−10M 8.02 2.05E−10M 1.18 (680 pM) (205 pM)

Full kinetic analysis was performed using 0 nM, 0.625 nM, 1.25 nM, 5 nM,and 10 nM of the analyte to determine K_(D). Chi square analysis wascarried out between the actual sensorgram and the sensorgram generatedfrom the BIAnalysis software to confirm the accuracy of the analysis;values of 1-2 were considered accurate and below 1 highly accurate.

TABLE 47 provides equilibrium dissociation constants based on fullkinetic experiments performed at multiple ligand concentrations. NB=nosignificant binding detected.

TABLE 47 VEGF VEGF HPTP-β HPTP-β Construct K_(D) chi sq K_(D) chi sqHC2:LC1 NB NB 1.21E−10M 0.123 (121 pM) Aflibercept 3.75E−11M 0.106 NB NB(37.5 pM) HC2-AFL:LC1 4.63E−11M 0.0875 2.19E−10M 0.0295 (46.3 pM) (219pM) HC2-BRO:LC1 2.18E−13M 0.452 1.54E−10M 0.0817 (218 fM) (154 pM)HC2-ABI:LC1 4.00E−14M 0.223 1.03E−12M 0.113 (40 fM) (1.03 pM) 

These results demonstrate that the multi-specific antibodiesHC2-AFL:LC1, HC2-BRO:LC1, HC2-ABI:LC1, HC2:LC1-AFL, HC2:LC1-BRO,HC2:LC1-ABI, HC2-AFL:LC1-AFL, HC2-BRO:LC1-BRO, and HC2-ABI:LC1-ABI bindto HPTP-β and VEGF with high affinity.

Example 19: A Hexavalent Antibody Comprising Antibody HC2:LC,Abicipar-Derived Sequences, and Brolucizumab-Derived Sequences

To generate a heavy chain with abicipar-derived VEGF-binding domainsadded at the C-terminus, an amino acid sequence is generated comprisingthe following appended amino acid sequences, from N-terminus toC-terminus:

1) Residues 1-467 of SEQ ID NO: 14 (heavy chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 244 (abicipar-derived sequence).

To generate a light chain with brolucizumab-derived VEGF-binding domainsadded at the C-terminus, an amino acid sequence is generated comprisingthe following appended amino acid sequences, from N-terminus toC-terminus:

1) SEQ ID NO: 17 (light chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 23 (brolucizumab-derived sequence).

The resulting polypeptides, SEQ ID NO: 231 and SEQ ID NO: 218, areco-expressed to provide a hexavalent antibody HC2-ABI:LC1-BRO shown inTABLE 48. Amino acids 1-19 of SEQ ID NO: 231 are the heavy chain signalpeptide (SEQ ID NO: 11). Amino acids 1-20 of SEQ ID NO: 218 are thelight chain signal peptide (SEQ ID NO: 12). In some embodiments, amature HC2-ABI:LC1-BRO does not comprise the signal peptides. Forexample, a mature HC2-ABI:LC1-BRO of the disclosure can comprise SEQ IDNO: 257 and SEQ ID NO: 258. The antibody is bispecific for targetmolecules, comprising a specificity for HPTP-β (VE-PTP) and VEGF. Theantibody is trispecific for target epitopes, comprising a specificityfor one HPTP-β (VE-PTP) epitope and for two VEGF epitopes.

TABLE 48 SEQ ID NO:  Name Amino acid sequence 231 SignalMGWTLVFLFLLSVTAGVHSEVQLVESGGGLVQPGGSLRLSC peptide-AASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATY HC2-YAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRD ABIYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSDLDKKLLEAARAGQDDEVRILMANGADVNARDSTGWTPLHLAAPWGHPEIVEVLLKNGADVNAADFQGWTPLHLAAAVGHLEIVEVLLKYGADVNAQ DKFGKTAFDISIDNGNEDLAEILQKAA 218Signal MVSSAQFLGLLLLCFQGTRCDVVMTQSPSFLSASVGDRVTIT peptide-CKASQHVGTAVAWYQQRPGKAPKLLIYWASTRHTGVPSRF LC1-SGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEI BROKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSEIVMTQSPSTLSASVGDRVIITCQASEIIHSWLAWYQQKPGKAPKLLIYLASTLASGVPSRFSGSGSGAEFTLTISSLQPDDFATYYCQNVYLASTNGANFGQGTKLTVLGGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCTASGFSLTDYYYMTWVRQAPGKGLEWVGFIDPDDDPYYATWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAGGDHNSGWGLDIWGQGTLVTVSS

Example 20: A Hexavalent Antibody Comprising Antibody HC2:LC,Aflibercept-Derived Sequences, and Brolucizumab-Derived Sequences

To generate a heavy chain with brolucizumab-derived VEGF-binding domainsadded at the C-terminus, an amino acid sequence is generated comprisingthe following appended amino acid sequences, from N-terminus toC-terminus:

1) SEQ ID NO: 14 (heavy chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 23 (brolucizumab-derived sequence).

To generate a light chain with aflibercept-derived VEGF-binding domainsadded at the C-terminus, an amino acid sequence is generated comprisingthe following appended amino acid sequences, from N-terminus toC-terminus:

1) SEQ ID NO: 17 (light chain of antibody HC2:LC1);

2) SEQ ID NO: 31 (linker peptide, underlined); and

3) SEQ ID NO: 22 (aflibercept-derived sequence).

The resulting polypeptides, SEQ ID NO: 150 and SEQ ID NO: 219, areco-expressed to provide a hexavalent antibody HC2-BRO:LC1-AFL shown inTABLE 49. Amino acids 1-19 of SEQ ID NO: 150 are the heavy chain signalpeptide (SEQ ID NO: 11). Amino acids 1-20 of SEQ ID NO: 219 are thelight chain signal peptide (SEQ ID NO: 12). In some embodiments, amature HC2-BRO:LC1-AFL does not comprise the signal peptides. Forexample, a mature HC2-BRO:LC1-AFL of the disclosure can comprise SEQ IDNO: 255 and SEQ ID NO: 259. The antibody is bispecific for targetmolecules, comprising a specificity for HPTP-β (VE-PTP) and VEGF. Theantibody is trispecific for target epitopes, comprising a specificityfor one HPTP-β (VE-PTP) epitope and for two VEGF epitopes.

TABLE 49 SEQ ID NO: Name Amino acid sequence 150 SignalMGWTLVFLFLLSVTAGVHSEVQLVESGGGLVQPGGSLRLSC peptide- AASGFTFNANAMNWVRQAPGKGLEWVGRIRTKSNNYATY HC2-YAGSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRD BROYYGSSAWITYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKGGGGSEIVMTQSPSTLSASVGDRVIITCQASEIIHSWLAWYQQKPGKAPKLLIYLASTLASGVPSRFSGSGSGAEFTLTISSLQPDDFATYYCQNVYLASTNGANFGQGTKLTVLGGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCTASGFSLTDYYYMTWVRQAPGKGLEWVGFIDPDDDPYYATWAKGRFTISRDNSKNTLYLQMNSLRAEDTA VYYCAGGDHNSGWGLDIWGQGTLVTVSS219 Signal MVSSAQFLGLLLLCFQGTRCDVVMTQSPSFLSASVGDRVTIT peptide-CKASQHVGTAVAWYQQRPGKAPKLLIYWASTRHTGVPSRF LC1-SGSGSGTEFTLTISSLQPEDFATYFCQQYSSYPFTFGGGTKLEIAFLKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSSDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSD QGLYTCAASSGLMTKKNSTFVRVHEK

Embodiments

The following non-limiting embodiments provide illustrative examples ofthe disclosure, but do not limit the scope of the disclosure.

Embodiment 1

A compound comprising: (a) a first domain, wherein the first domainmodulates a phosphatase, wherein the phosphatase modulates Tie2; and (b)a second domain that specifically binds a receptor tyrosine kinaseagonist.

Embodiment 2

The compound of embodiment 1, wherein the compound is an antibody.

Embodiment 3

The compound of any one of embodiments 1-2, wherein the compound is amultispecific antibody.

Embodiment 4

The compound of any one of embodiments 1-3, wherein the compound is atetravalent antibody.

Embodiment 5

The compound of any one of embodiments 1-3, wherein the compound is ahexavalent antibody.

Embodiment 6

The compound of any one of embodiments 1-5, wherein the compound is abispecific antibody.

Embodiment 7

The compound of any one of embodiments 1-4 and 6, wherein the compoundis a tetravalent bispecific antibody.

Embodiment 8

The compound of any one of embodiments 1-3 and 5-6, wherein the compoundis a hexavalent bispecific antibody.

Embodiment 9

The compound of any one of embodiments 1-8, wherein the compoundinhibits the phosphatase that modulates Tie2.

Embodiment 10

The compound of any one of embodiments 1-9, wherein the compoundinhibits HPTP-β.

Embodiment 11

The compound of any one of embodiments 1-10, wherein the compoundinhibits VE-PTP.

Embodiment 12

The compound of any one of embodiments 1-11, wherein the compoundactivates Tie2.

Embodiment 13

The compound of any one of embodiments 1-12, wherein the compoundinhibits the receptor tyrosine kinase agonist.

Embodiment 14

The compound of any one of embodiments 1-13, wherein the compoundinhibits VEGF receptor signaling.

Embodiment 15

The compound of any one of embodiments 1-14, wherein the compoundinhibits a VEGF.

Embodiment 16

The compound of any one of embodiments 1-15, wherein the compoundinhibits VEGF-A.

Embodiment 17

The compound of any one of embodiments 1-16, wherein the compoundinhibits the phosphatase that modulates Tie2, and inhibits the receptortyrosine kinase agonist.

Embodiment 18

The compound of any one of embodiments 1-17, wherein the phosphatase isHPTP-β, and the receptor tyrosine kinase agonist is a VEGF.

Embodiment 19

The compound of any one of embodiments 1-18, wherein the phosphatase isHPTP-β, and the receptor tyrosine kinase agonist is VEGF-A.

Embodiment 20

The compound of any one of embodiments 1-19, wherein the compoundactivates Tie2, and the receptor tyrosine kinase agonist is a VEGF.

Embodiment 21

The compound of any one of embodiments 1-20, wherein the compoundactivates Tie2, and the receptor tyrosine kinase agonist is VEGF-A.

Embodiment 22

The compound of any one of embodiments 1-21, wherein the phosphatasethat modulates Tie2 signaling is a protein tyrosine phosphatase.

Embodiment 23

The compound of any one of embodiments 1-22, wherein the phosphatasethat modulates Tie2 signaling is a receptor-like protein tyrosinephosphatase.

Embodiment 24

The compound of any one of embodiments 1-23, wherein the phosphatasethat modulates Tie2 signaling is HPTP-β.

Embodiment 25

The compound of any one of embodiments 1-23, wherein the phosphatasethat modulates Tie2 signaling is VE-PTP.

Embodiment 26

The compound of any one of embodiments 1-25, wherein the receptortyrosine kinase agonist is a growth factor.

Embodiment 27

The compound of any one of embodiments 1-26, wherein the receptortyrosine kinase agonist is a cysteine-knot growth factor superfamilymember.

Embodiment 28

The compound of any one of embodiments 1-27, wherein the receptortyrosine kinase agonist is a PDGF family member.

Embodiment 29

The compound of any one of embodiments 1-28, wherein the receptortyrosine kinase agonist is a pro-angiogenic factor.

Embodiment 30

The compound of any one of embodiments 1-29, wherein the receptortyrosine kinase agonist is a VEGF receptor agonist.

Embodiment 31

The compound of any one of embodiments 1-30, wherein the receptortyrosine kinase agonist is a VEGF.

Embodiment 32

The compound of any one of embodiments 1-31, wherein the receptortyrosine kinase agonist is VEGF-A.

Embodiment 33

The compound of any one of embodiments 1-32, wherein the first domainbinds to HPTP-β.

Embodiment 34

The compound of any one of embodiments 1-33, wherein the first domainbinds to VE-PTP.

Embodiment 35

The compound of any one of embodiments 1-34, wherein the first domainbinds to an extracellular domain of HPTP-β.

Embodiment 36

The compound of any one of embodiments 1-34, wherein the first domainbinds to a first FN3 repeat of an extracellular domain of HPTP-β.

Embodiment 37

The compound of any one of embodiments 1-36, wherein the second domainbinds to a VEGF.

Embodiment 38

The compound of any one of embodiments 1-37, wherein the second domainbinds to VEGF-A.

Embodiment 39

The compound of any one of embodiments 1-38, wherein the first domainbinds to HPTP-β, and the receptor tyrosine kinase agonist is a VEGF.

Embodiment 40

The compound of any one of embodiments 1-39, wherein the first domainbinds to HPTP-β, and the receptor tyrosine kinase agonist is VEGF-A.

Embodiment 41

The compound of any one of embodiments 1-40, wherein the first domaincomprises a sequence that is at least 80% identical to any one of SEQ IDNOS: 76-98.

Embodiment 42

The compound of any one of embodiments 1-41, wherein the first domaincomprises a sequence that is at least 80% identical to SEQ ID NO: 76,SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, or SEQ ID NO: 80.

Embodiment 43

The compound of any one of embodiments 1-42, wherein the first domaincomprises a sequence that is at least 80% identical to SEQ ID NO: 81,SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO:86, or SEQ ID NO: 87.

Embodiment 44

The compound of any one of embodiments 1-43, wherein the first domaincomprises a sequence that is at least 80% identical to SEQ ID NO: 88,SEQ ID NO: 89, or SEQ ID NO: 90.

Embodiment 45

The compound of any one of embodiments 1-44, wherein the first domaincomprises a sequence that is at least 80% identical to SEQ ID NO: 91,SEQ ID NO: 92, or SEQ ID NO: 93.

Embodiment 46

The compound of any one of embodiments 1-45, wherein the first domaincomprises a sequence that is at least 80% identical to SEQ ID NO: 94,SEQ ID NO: 95, or SEQ ID NO: 96.

Embodiment 47

The compound of any one of embodiments 1-46, wherein the first domaincomprises a sequence that is at least 80% identical to SEQ ID NO: 97 orSEQ ID NO: 98.

Embodiment 48

The compound of any one of embodiments 1-47, wherein the first domaincomprises: (a) a sequence that is at least 80% identical to SEQ ID NO:76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, or SEQ ID NO: 80; (b) asequence that is at least 80% identical to SEQ ID NO: 81, SEQ ID NO: 82,SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, or SEQ IDNO: 87; (c) a sequence that is at least 80% identical to SEQ ID NO: 88,SEQ ID NO: 89, or SEQ ID NO: 90; (d) a sequence that is at least 80%identical to SEQ ID NO: 91, SEQ ID NO: 92, or SEQ ID NO: 93; (e) asequence that is at least 80% identical to SEQ ID NO: 94, SEQ ID NO: 95,or SEQ ID NO: 96; and (f) a sequence that is at least 80% identical toSEQ ID NO: 97 or SEQ ID NO: 98.

Embodiment 49

The compound of any one of embodiments 1-48, wherein the second domaincomprises a sequence that is at least 80% identical to SEQ ID NO: 22,SEQ ID NO: 23, SEQ ID NO: 28, or SEQ ID NO: 244.

Embodiment 50

The compound of any one of embodiments 1-49, wherein the second domaincomprises a sequence that is at least 80% identical to any one of SEQ IDNOS: 99-146.

Embodiment 51

The compound of any one of embodiments 1-50, wherein the second domaincomprises a sequence that is at least 80% identical to SEQ ID NO: 99,SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ IDNO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108,SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, or SEQID NO: 113.

Embodiment 52

The compound of any one of embodiments 1-51, wherein the second domaincomprises a sequence that is at least 80% identical to SEQ ID NO: 114,SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ IDNO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, or SEQ ID NO:123.

Embodiment 53

The compound of any one of embodiments 1-52, wherein the second domaincomprises a sequence that is at least 80% identical to SEQ ID NO: 124,SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ IDNO: 129, SEQ ID NO: 130, or SEQ ID NO: 131.

Embodiment 54

The compound of any one of embodiments 1-53, wherein the second domaincomprises a sequence that is at least 80% identical to SEQ ID NO: 132,SEQ ID NO: 133, SEQ ID NO: 134, SEQ ID NO: 135, SEQ ID NO: 136, or SEQID NO: 137.

Embodiment 55

The compound of any one of embodiments 1-54, wherein the second domaincomprises a sequence that is at least 80% identical to SEQ ID NO: 138,SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 141, SEQ ID NO: 142, or SEQID NO: 143.

Embodiment 56

The compound of any one of embodiments 1-55, wherein the second domaincomprises a sequence that is at least 80% identical to SEQ ID NO: 144,SEQ ID NO: 145, or SEQ ID NO: 146.

Embodiment 57

The compound of any one of embodiments 1-56, wherein the second domaincomprises: (a) a sequence that is at least 80% identical to SEQ ID NO:99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO:108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, orSEQ ID NO: 113; (b) a sequence that is at least 80% identical to SEQ IDNO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118,SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, or SEQID NO: 123; (c) a sequence that is at least 80% identical to SEQ ID NO:124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQID NO: 129, SEQ ID NO: 130, or SEQ ID NO: 131; (d) a sequence that is atleast 80% identical to SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO: 134,SEQ ID NO: 135, SEQ ID NO: 136, or SEQ ID NO: 137; (e) a sequence thatis at least 80% identical to SEQ ID NO: 138, SEQ ID NO: 139, SEQ ID NO:140, SEQ ID NO: 141, SEQ ID NO: 142, or SEQ ID NO: 143; and (f) asequence that is at least 80% identical to SEQ ID NO: 144, SEQ ID NO:145, or SEQ ID NO: 146.

Embodiment 58

The compound of any one of embodiments 1-57, wherein the second domaincomprises a sequence that is at least 80% identical to SEQ ID NO: 147,SEQ ID NO: 148, or any one of SEQ ID NOS: 233-242.

Embodiment 59

The compound of any one of embodiments 1-58, wherein: (a) the firstdomain comprises a sequence that is at least 80% identical to any one ofSEQ ID NOS: 76-98; and (b) the second domain comprises a sequence thatis at least 80% identical to any one of SEQ ID NOS: 99-148 or any one ofSEQ ID NOS: 233-242.

Embodiment 60

The compound of any one of embodiments 1-59, wherein: (a) the firstdomain comprises: (i) a sequence that is at least 80% identical to SEQID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, or SEQ ID NO:80; (ii) a sequence that is at least 80% identical to SEQ ID NO: 81, SEQID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86,or SEQ ID NO: 87; (iii) a sequence that is at least 80% identical to SEQID NO: 88, SEQ ID NO: 89, or SEQ ID NO: 90; (iv) a sequence that is atleast 80% identical to SEQ ID NO: 91, SEQ ID NO: 92, or SEQ ID NO: 93;(v) a sequence that is at least 80% identical to SEQ ID NO: 94, SEQ IDNO: 95, or SEQ ID NO: 96; and (vi) a sequence that is at least 80%identical to SEQ ID NO: 97 or SEQ ID NO: 98; and (b) the second domaincomprises: (i) a sequence that is at least 80% identical to SEQ ID NO:99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO:108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, orSEQ ID NO: 113; (ii) a sequence that is at least 80% identical to SEQ IDNO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118,SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, or SEQID NO: 123; (iii) a sequence that is at least 80% identical to SEQ IDNO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128,SEQ ID NO: 129, SEQ ID NO: 130, or SEQ ID NO: 131; (iv) a sequence thatis at least 80% identical to SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO:134, SEQ ID NO: 135, SEQ ID NO: 136, or SEQ ID NO: 137; (v) a sequencethat is at least 80% identical to SEQ ID NO: 138, SEQ ID NO: 139, SEQ IDNO: 140, SEQ ID NO: 141, SEQ ID NO: 142, or SEQ ID NO: 143; and (vi) asequence that is at least 80% identical to SEQ ID NO: 144, SEQ ID NO:145, or SEQ ID NO: 146.

Embodiment 61

The compound of any one of embodiments 1-60, wherein: (a) the firstdomain comprises a sequence that is at least 80% identical to SEQ ID NO:76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, or SEQ ID NO: 80; and(b) the second domain comprises a sequence that is at least 80%identical to SEQ ID NO: 22, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO:244, or any one of SEQ ID NOS: 233-242.

Embodiment 62

The compound of any one of embodiments 1-61, wherein: (a) the firstdomain comprises a sequence that is at least 80% identical to SEQ ID NO:81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ IDNO: 86, or SEQ ID NO: 87; and (b) the second domain comprises a sequencethat is at least 80% identical to SEQ ID NO: 22, SEQ ID NO: 147, SEQ IDNO: 148, SEQ ID NO: 244, or any one of SEQ ID NOS: 233-242.

Embodiment 63

The compound of any one of embodiments 1-62, wherein: (a) the firstdomain comprises a sequence that is at least 80% identical to SEQ ID NO:88, SEQ ID NO: 89, or SEQ ID NO: 90; and (b) the second domain comprisesa sequence that is at least 80% identical to SEQ ID NO: 22, SEQ ID NO:147, SEQ ID NO: 148, SEQ ID NO: 244, or any one of SEQ ID NOS: 233-242.

Embodiment 64

The compound of any one of embodiments 1-63, wherein: (a) the firstdomain comprises a sequence that is at least 80% identical to SEQ ID NO:91, SEQ ID NO: 92, or SEQ ID NO: 93; and (b) the second domain comprisesa sequence that is at least 80% identical to SEQ ID NO: 22, SEQ ID NO:147, SEQ ID NO: 148, SEQ ID NO: 244, or any one of SEQ ID NOS: 233-242.

Embodiment 65

The compound of any one of embodiments 1-64, wherein: (a) the firstdomain comprises a sequence that is at least 80% identical to SEQ ID NO:94, SEQ ID NO: 95, or SEQ ID NO: 96; and (b) the second domain comprisesa sequence that is at least 80% identical to SEQ ID NO: 22, SEQ ID NO:147, SEQ ID NO: 148, SEQ ID NO: 244, or any one of SEQ ID NOS: 233-242.

Embodiment 66

The compound of any one of embodiments 1-65, wherein: (a) the firstdomain comprises a sequence that is at least 80% identical to SEQ ID NO:97 or SEQ ID NO: 98; and (b) the second domain comprises a sequence thatis at least 80% identical to SEQ ID NO: 22, SEQ ID NO: 147, SEQ ID NO:148, SEQ ID NO: 244, or any one of SEQ ID NOS: 233-242.

Embodiment 67

The compound of any one of embodiments 1-66, wherein: (a) the firstdomain comprises: (i) a sequence that is at least 80% identical to SEQID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, or SEQ ID NO:80; (ii) a sequence that is at least 80% identical to SEQ ID NO: 81, SEQID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86,or SEQ ID NO: 87; (iii) a sequence that is at least 80% identical to SEQID NO: 88, SEQ ID NO: 89, or SEQ ID NO: 90; (iv) a sequence that is atleast 80% identical to SEQ ID NO: 91, SEQ ID NO: 92, or SEQ ID NO: 93;(v) a sequence that is at least 80% identical to SEQ ID NO: 94, SEQ IDNO: 95, or SEQ ID NO: 96; and (vi) a sequence that is at least 80%identical to SEQ ID NO: 97 or SEQ ID NO: 98; and (b) the second domaincomprises a sequence that is at least 80% identical to SEQ ID NO: 22,SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 244, or any one of SEQ IDNOS: 233-242.

Embodiment 68

The compound of any one of embodiments 1-67, wherein the compoundcomprises: (a) a heavy chain sequence that is at least 80% identical toSEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 151, or SEQ ID NO: 231; and(b) a light chain sequence that is at least 80% identical to SEQ ID NO:17, SEQ ID NO: 18, SEQ ID NO: 19, or SEQ ID NO: 20.

Embodiment 69

The compound of any one of embodiments 1-4, 6-7, and 9-68, wherein thecompound is a tetravalent bispecific antibody comprising: (a) a sequencethat is at least 80% identical to SEQ ID NO: 22, SEQ ID NO: 23, SEQ IDNO: 28, or SEQ ID NO: 244; (b) a sequence that is at least 80% identicalto SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, or SEQ ID NO: 16; (c) asequence that is at least 80% identical to any one of SEQ ID NOS: 31-75;and (d) a sequence that is at least 80% identical to SEQ ID NO: 17, SEQID NO: 18, SEQ ID NO: 19, or SEQ ID NO: 20.

Embodiment 70

The compound of any one of embodiments 1-4, 6-7 and 9-69, wherein thecompound is a tetravalent bispecific antibody comprising: (a) a firstchain, wherein the first chain comprises a linker, wherein the linkercomprises a sequence that is at least 80% identical to any one of SEQ IDNOS: 31-75, wherein the linker comprises a N-terminus and a C-terminus,wherein the N-terminus of the linker is attached to a C terminus of asequence that is at least 80% identical to SEQ ID NO: 13, SEQ ID NO: 14,SEQ ID NO: 15, or SEQ ID NO: 16, and the C-terminus of the linker isattached to a N-terminus of a sequence that is at least 80% identical toSEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 28, or SEQ ID NO: 244; and (b)a second chain, wherein the second chain comprises a sequence that is atleast 80% identical to SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, orSEQ ID NO: 20.

Embodiment 71

The compound of any one of embodiments 1-4, 6-7, 9-67, and 69, whereinthe compound is a tetravalent bispecific antibody comprising: (a) afirst chain, wherein the first chain comprises a sequence that is atleast 80% identical to SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, orSEQ ID NO: 16; and (b) a second chain, wherein the second chaincomprises a linker, wherein the linker comprises a sequence that is atleast 80% identical to any one of SEQ ID NOS: 31-75, wherein the linkercomprises a N-terminus and a C-terminus, wherein the N-terminus of thelinker is attached to a C terminus of a sequence that is at least 80%identical to SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, or SEQ ID NO:20, and the C-terminus of the linker is attached to a N-terminus of asequence that is at least 80% identical to SEQ ID NO: 22, SEQ ID NO: 23,SEQ ID NO: 28, or SEQ ID NO: 244.

Embodiment 72

The compound of any one of embodiments 1-3, 5-6, and 8-67, wherein thecompound is a hexavalent bispecific antibody comprising: (a) a sequencethat is at least 80% identical to SEQ ID NO: 22, SEQ ID NO: 23, SEQ IDNO: 28, or SEQ ID NO: 244; (b) a sequence that is at least 80% identicalto SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, or SEQ ID NO: 16; (c) asequence that is at least 80% identical to any one of SEQ ID NOS: 31-75;and (d) a sequence that is at least 80% identical to SEQ ID NO: 17, SEQID NO: 18, SEQ ID NO: 19, or SEQ ID NO: 20.

Embodiment 73

The compound of any one of embodiments 1-3, 5-6, 8-67, and 72, whereinthe compound is a hexavalent bispecific antibody comprising: (a) a firstchain, wherein the first chain comprises a linker, wherein the linkercomprises a sequence that is at least 80% identical to any one of SEQ IDNOS: 31-75, wherein the linker comprises a N-terminus and a C-terminus,wherein the N-terminus of the linker is attached to a C terminus of asequence that is at least 80% identical to SEQ ID NO: 13, SEQ ID NO: 14,SEQ ID NO: 15, or SEQ ID NO: 16, and the C-terminus of the linker isattached to a N-terminus of a sequence that is at least 80% identical toSEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 28, or SEQ ID NO: 244; and (b)a second chain, wherein the second chain comprises a linker, wherein thelinker comprises a sequence that is at least 80% identical to any one ofSEQ ID NOS: 31-75, wherein the linker comprises a N-terminus and aC-terminus, wherein the N-terminus of the linker is attached to a Cterminus of a sequence that is at least 80% identical to SEQ ID NO: 17,SEQ ID NO: 18, SEQ ID NO: 19, or SEQ ID NO: 20, and the C-terminus ofthe linker is attached to a N-terminus of a sequence that is at least80% identical to SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 28, or SEQ IDNO: 244.

Embodiment 74

The compound of any one of embodiments 1-67, wherein the compoundcomprises: (a) a heavy chain sequence that is at least 80% identical toSEQ ID NO: 245 or any one of SEQ ID NOS: 13-16; and (b) a light chainsequence that is at least 80% identical to SEQ ID NO: 218, SEQ ID NO:219, SEQ ID NO: 232, or SEQ ID NO: 243.

Embodiment 75

The compound of any one of embodiments 1-67, wherein the compoundcomprises: (a) a heavy chain sequence that is at least 80% identical toSEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 151, or SEQ ID NO: 231; and(b) a light chain sequence that is at least 80% identical to SEQ ID NO:218, SEQ ID NO: 219, SEQ ID NO: 232, or SEQ ID NO: 243.

Embodiment 76

The compound of any one of embodiments 1-75, wherein a binding affinity(K_(D)) of the compound to HPTP-β is about 30 fM to about 70 nM.

Embodiment 77

The compound of any one of embodiments 1-76, wherein a binding affinity(K_(D)) of the compound to the VEGF is about 30 fM to about 70 nM.

Embodiment 78

The compound of any one of embodiments 1-75, wherein a binding affinity(K_(D)) of the compound to HPTP-β is about 30 fM to about 70 nM, and abinding affinity (K_(D)) of the compound to VEGF is about 30 fM to about70 nM.

Embodiment 79

A method of treating a condition in a subject in need thereof, themethod comprising administering to the subject atherapeutically-effective amount of the compound of any one ofembodiments 1-78.

Embodiment 80

The method of embodiment 79, wherein the condition is an ocularcondition.

Embodiment 81

The method of any one of embodiments 79-80, wherein the condition isdiabetic retinopathy.

Embodiment 82

The method of any one of embodiments 79-80, wherein the condition isneovascularization.

Embodiment 83

The method of any one of embodiments 79-80, wherein the condition isvascular leak.

Embodiment 84

The method of any one of embodiments 79-80, wherein the condition isincreased intraocular pressure.

Embodiment 85

The method of any one of embodiments 79-80, wherein the condition isocular edema.

Embodiment 86

The method of any one of embodiments 79-80 and 85, wherein the conditionis diabetic macular edema.

Embodiment 87

The method of any one of embodiments 79-80, wherein the condition isocular hypertension.

Embodiment 88

The method of any one of embodiments 79-80, wherein the condition isocular inflammation.

Embodiment 89

The method of any one of embodiments 79-80, wherein the condition isglaucoma.

Embodiment 90

The method of any one of embodiments 79-89, wherein the administrationis to an eye of the subject.

Embodiment 91

The method of any one of embodiments 79-90, wherein the administrationis intravitreal.

Embodiment 92

The method of any one of embodiments 79-89, wherein the administrationis subcutaneous.

Embodiment 93

The method of any one of embodiments 79-90, wherein the administrationis topical.

Embodiment 94

The method of any one of embodiments 79-93, wherein the subject ishuman.

Embodiment 95

The method of any one of embodiments 79-94, wherein thetherapeutically-effective amount is from about 0.25 mg to about 200 mg.

Embodiment 96

The method of any one of embodiments 79-95, wherein thetherapeutically-effective amount is from about 1 mg/kg to about 10mg/kg.

Embodiment 97

The method of any one of embodiments 79-95, wherein thetherapeutically-effective amount is from about 1 mg to about 50 mg.

Embodiment 98

The method of any one of embodiments 79-95, wherein thetherapeutically-effective amount is from about 50 mg to about 200 mg.

Embodiment 99

The compound of any one of embodiments 1-67, wherein the compoundcomprises: (a) a heavy chain sequence that is at least 80% identical toSEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, or SEQ ID NO: 257; and(b) a light chain sequence that is at least 80% identical to SEQ ID NO:250, SEQ ID NO: 251, SEQ ID NO: 252, or SEQ ID NO: 253.

Embodiment 100

The compound of any one of embodiments 1-4, 6-7, and 9-68, wherein thecompound is a tetravalent bispecific antibody comprising: (a) a sequencethat is at least 80% identical to SEQ ID NO: 22, SEQ ID NO: 23, SEQ IDNO: 28, or SEQ ID NO: 244; (b) a sequence that is at least 80% identicalto SEQ ID NO: 246, SEQ ID NO: 247, SEQ ID NO: 248, or SEQ ID NO: 249;(c) a sequence that is at least 80% identical to any one of SEQ ID NOS:31-75; and (d) a sequence that is at least 80% identical to SEQ ID NO:250, SEQ ID NO: 251, SEQ ID NO: 252, or SEQ ID NO: 253.

Embodiment 101

The compound of any one of embodiments 1-4, 6-7 and 9-69, wherein thecompound is a tetravalent bispecific antibody comprising: (a) a firstchain, wherein the first chain comprises a linker, wherein the linkercomprises a sequence that is at least 80% identical to any one of SEQ IDNOS: 31-75, wherein the linker comprises a N-terminus and a C-terminus,wherein the N-terminus of the linker is attached to a C terminus of asequence that is at least 80% identical to SEQ ID NO: 246, SEQ ID NO:247, SEQ ID NO: 248, or SEQ ID NO: 249, and the C-terminus of the linkeris attached to a N-terminus of a sequence that is at least 80% identicalto SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 28, or SEQ ID NO: 244; and(b) a second chain, wherein the second chain comprises a sequence thatis at least 80% identical to SEQ ID NO: 250, SEQ ID NO: 251, SEQ ID NO:252, or SEQ ID NO: 253.

Embodiment 102

The compound of any one of embodiments 1-4, 6-7, 9-67, and 69, whereinthe compound is a tetravalent bispecific antibody comprising: (a) afirst chain, wherein the first chain comprises a sequence that is atleast 80% identical to SEQ ID NO: 246, SEQ ID NO: 247, SEQ ID NO: 248,or SEQ ID NO: 249; and (b) a second chain, wherein the second chaincomprises a linker, wherein the linker comprises a sequence that is atleast 80% identical to any one of SEQ ID NOS: 31-75, wherein the linkercomprises a N-terminus and a C-terminus, wherein the N-terminus of thelinker is attached to a C terminus of a sequence that is at least 80%identical to SEQ ID NO: 250, SEQ ID NO: 251, SEQ ID NO: 252, or SEQ IDNO: 253, and the C-terminus of the linker is attached to a N-terminus ofa sequence that is at least 80% identical to SEQ ID NO: 22, SEQ ID NO:23, SEQ ID NO: 28, or SEQ ID NO: 244.

Embodiment 103

The compound of any one of embodiments 1-3, 5-6, and 8-67, wherein thecompound is a hexavalent bispecific antibody comprising: (a) a sequencethat is at least 80% identical to SEQ ID NO: 22, SEQ ID NO: 23, SEQ IDNO: 28, or SEQ ID NO: 244; (b) a sequence that is at least 80% identicalto SEQ ID NO: 246, SEQ ID NO: 247, SEQ ID NO: 248, or SEQ ID NO: 249;(c) a sequence that is at least 80% identical to any one of SEQ ID NOS:31-75; and (d) a sequence that is at least 80% identical to SEQ ID NO:250, SEQ ID NO: 251, SEQ ID NO: 252, or SEQ ID NO: 253.

Embodiment 104

The compound of any one of embodiments 1-3, 5-6, 8-67, and 72, whereinthe compound is a hexavalent bispecific antibody comprising: (a) a firstchain, wherein the first chain comprises a linker, wherein the linkercomprises a sequence that is at least 80% identical to any one of SEQ IDNOS: 31-75, wherein the linker comprises a N-terminus and a C-terminus,wherein the N-terminus of the linker is attached to a C terminus of asequence that is at least 80% identical to SEQ ID NO: 246, SEQ ID NO:247, SEQ ID NO: 248, or SEQ ID NO: 249, and the C-terminus of the linkeris attached to a N-terminus of a sequence that is at least 80% identicalto SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 28, or SEQ ID NO: 244; and(b) a second chain, wherein the second chain comprises a linker, whereinthe linker comprises a sequence that is at least 80% identical to anyone of SEQ ID NOS: 31-75, wherein the linker comprises a N-terminus anda C-terminus, wherein the N-terminus of the linker is attached to a Cterminus of a sequence that is at least 80% identical to SEQ ID NO: 250,SEQ ID NO: 251, SEQ ID NO: 252, or SEQ ID NO: 253, and the C-terminus ofthe linker is attached to a N-terminus of a sequence that is at least80% identical to SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 28, or SEQ IDNO: 244.

Embodiment 105

The compound of any one of embodiments 1-67, wherein the compoundcomprises: (a) a heavy chain sequence that is at least 80% identical toSEQ ID NO: 262 or any one of SEQ ID NOS: 246-249; and (b) a light chainsequence that is at least 80% identical to SEQ ID NO: 258, SEQ ID NO:259, SEQ ID NO: 261, or SEQ ID NO: 260.

Embodiment 106

The compound of any one of embodiments 1-3, 5-6, and 8-67, wherein thecompound comprises: (a) a heavy chain sequence that is at least 80%identical to SEQ ID NO: 254, SEQ ID NO: 255, SEQ ID NO: 256, or SEQ IDNO: 257; and (b) a light chain sequence that is at least 80% identicalto SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO: 261, or SEQ ID NO: 260.

What is claimed is:
 1. A method of treating an ocular condition in asubject in need thereof, the method comprising administering to thesubject a therapeutically effective amount of an antibody comprising:(a) a first domain that activates Tie2; and (b) a second domain thatspecifically binds a VEGF and inhibits VEGF function.
 2. The method ofclaim 1, wherein the ocular condition is diabetic retinopathy.
 3. Themethod of claim 1, wherein the ocular condition is neovascularization.4. The method of claim 1, wherein the ocular condition is vascular leak.5. The method of claim 1, wherein the ocular condition is increasedintraocular pressure.
 6. The method of claim 1, wherein the ocularcondition is ocular edema.
 7. The method of claim 1, wherein the ocularcondition is diabetic macular edema.
 8. The method of claim 1, whereinthe ocular condition is wet age-related macular degeneration.
 9. Themethod of claim 1, wherein the ocular condition is ocular inflammation.10. The method of claim 1, wherein the ocular condition is retinal veinocclusion.
 11. The method of claim 1, wherein the administration is toan eye of the subject.
 12. The method of claim 1, wherein theadministration is intravitreal.
 13. The method of claim 1, wherein theadministration is subcutaneous.
 14. The method of claim 1, wherein theadministration is systemic.
 15. The method of claim 1, wherein thesubject is a human.
 16. The method of claim 1, wherein thetherapeutically effective amount is from about 0.25 mg to about 200 mg.17. The method of claim 1, wherein the therapeutically effective amountis from about 1 mg/kg to about 10 mg/kg.
 18. The method of claim 1,wherein the therapeutically effective amount is from about 1 mg to about50 mg.
 19. The method of claim 1, wherein the therapeutically effectiveamount is from about 50 mg to about 200 mg.